Compounds and compositions for use in phototherapy and in treatment of ocular neovascular disease and cancers

ABSTRACT

The invention relates generally to anti-angiogenesis agents and related methods of using to anti-angiogenesis agents for biomedical applications including direct monotherapy and combination therapy for treatment of an angiogenesis related condition. In an embodiment, the invention provides a class of opioid compounds and structurally related opioid derivatives exhibiting anti-VEGF activity for use in therapeutic procedures, including phototherapy. Opioid compounds and structurally related opioid derivatives of the invention may be administered alone or in combination with administration of a phototherapy agent and/or other therapeutic agent.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a U.S. National Stage Application under 35 U.S.C.§371 of International Application No. PCT/US10/042218, filed Jul. 16,2010, which claims the benefit of U.S. Provisional Application No.61/226,032 filed Jul. 16, 2009 and U.S. Provisional Application No.61/238,358 filed Aug. 31, 2009, U.S. Provisional Application No.61/226,015, filed on Jul. 16, 2009, and U.S. Provisional Application No.61/286,877, filed on Dec. 16, 2009, all of which are incorporated byreference to the extent not inconsistent herewith.

BACKGROUND

Optical agents play a central role in a large number of in vivo, invitro and ex vivo clinical procedures including important diagnostic andtherapeutic procedures. Photodiagnostic and phototherapy agents, forexample, include a class of molecules capable of absorbing, emitting, orscattering electromagnetic radiation applied to a biological material,particularly in the visible and near infrared regions of theelectromagnetic spectrum. This property of optical agents is used in arange of biomedical applications for visualizing, imaging or otherwisecharacterizing biological materials and/or achieving a desiredtherapeutic outcome. Recent developments in targeted administration anddelivery of optical agents, and advanced systems and methods forapplying and detecting electromagnetic radiation in biologicalenvironments, has considerably expanded the applicability andeffectiveness of optical agents for many clinical applications.

Important applications of optical agents that absorb and/or emit in thevisible and near-infrared (NIR) region of the electromagnetic spectruminclude their use in biomedical imaging and visualization. Imaging andvisualization using such optical agents has potential to provide a lessinvasive and safer imaging technology, as compared to X-ray, and otherwidely used nuclear medicine technologies. Applications of opticalimaging for diagnosis and monitoring of the onset, progression andtreatment of various disease conditions, including cancer, are wellestablished. (See, e.g., D. A. Benaron and D. K. Stevenson, Opticaltime-of-flight and absorbance imaging of biologic media, Science, 1993,259, pp. 1463-1466; R. F. Potter (Series Editor), Medical opticaltomography: functional imaging and monitoring, SPIE Optical EngineeringPress, Bellingham, 1993; G. J. Tearney et al., In vivo endoscopicoptical biopsy with optical coherence tomography, Science, 1997, 276,pp. 2037-2039; B. J. Tromberg et al., Non-invasive measurements ofbreast tissue optical properties using frequency-domain photonmigration, Phil. Trans. Royal Society London B, 1997, 352, pp. 661-668;S. Fantini et al., Assessment of the size, position, and opticalproperties of breast tumors in vivo by noninvasive optical methods,Appl. Opt., 1998, 37, pp. 1982-1989; A. Pelegrin et al.,Photoimmunodiagnosis with antibody-fluorescein conjugates: in vitro andin vivo preclinical studies, J. Cell Pharmacol., 1992, 3, pp. 141-145).

In addition to their important role in biomedical imaging andvisualization, optical agents capable of absorption in the visible andNIR regions have also been extensively developed for clinicalapplications for phototherapy. The benefits of phototherapy usingoptical agents are widely acknowledged as this technique has thepotential to provide efficacy comparable to radiotherapy, while entirelyavoiding exposure of non-target organs and tissue to harmful ionizingradiation. Photodynamic therapy (PDT), in particular, has been usedeffectively for localized superficial or endoluminal malignant andpremalignant conditions. The clinical efficacy of PDT has also beendemonstrated for the treatment of various other diseases, injuries, anddisorders, including cardiovascular disorders such as atherosclerosisand vascular restenosis, inflammatory diseases, ophthalmic diseases suchas age related macular degeneration and dermatological diseases.Visudyne® and Photofrin®, for example, are two optical agents that havebeen developed and approved by the FDA for the treatment of maculardegeneration of the eye and for ablation of several types of tumors,respectively. (See, e.g., Schmidt-Drfurth, U.; Bringruber, R.; Hasan, T.Phototherapy in ocular vascular disease. IEEE Journal of Selected Topicsin Quantum Electronics 1996, 2, 988-996; Mlkvy, P.; Messmann, H.;Regula, J.; Conio, M.; Pauer, M.; Millson, C. E.; MacRobert, A. J.;Brown, S. G. Phototherapy for gastrointestinal tumors using threephotosensitizers—ALA induced PPIX, Photofrin, and MTHPC. A pilot study.Neoplasma 1998, 45, 157-161; Grosjean, P.; Wagieres, G.; Fontolliet, C.;Van Den Bergh, H.; Monnier, P. Clinical phototherapy for superficialcancer in the esophagus and the bronchi: 514 nm compared with 630 nmlight irradiation after sensitization with Photofrin II. British Journalof Cancer 1998, 77, 1989-1955; Mitton, D.; Ackroyd, R. Phototherapy ofBarrett's oesophagus and oesophageal carcinoma—how I do it.Photodiagnostics and Phototherapy 2006, 3, 96-98; and Li, L.; Luo, R.;Liao, W.; Zhang, M.; Luo, Y.; Miao, J. Clinical study of photofrinphototherapy for the treatment of relapse nasopharyngeal carcinoma.Photodiagnostics and Phototherapy 2006, 3, 266-271).

Phototherapy is carried out by administration, and preferably targeteddelivery, of a photosensitizer to a target tissue (e.g., tumor, lesion,organ etc.) followed by photoactivation of the photosensitizer byabsorption of applied electromagnetic radiation, typically provided by alaser light source. Phototherapy targets include tumor cells, tumormicrovaculature, inflammatory cells, immune host cells and neovascularendothelium cells. The applied electromagnetic radiation excites thephotosensitizer, resulting in formation of reactive species capable ofinitiating a cascade of cellular and molecular events eventuallyresulting in selective target tissue destruction.

Photosensitizers may operate via two different major pathways,classified as Types 1 and 2. The Type 1 mechanism proceeds via atwo-step process involving activation of the photosensitizer by appliedelectromagnetic radiation followed either by direct transfer of theenergy from the excited state of the photosensitizer to the tissue, orthough the interaction of reactive intermediates (e.g., radicals, ions,nitrene, carbene etc.) derived from the excited photosensitizer with thetarget tissue, resulting in tissue damage. The Type 1 mechanism can berepresented by the following sequence of reactions:Step 1: PHOTOSENSITIZER+hv→(PHOTOSENSITIZER)*  (1)Step 2: (PHOTOSENSITIZER)*+TISSUE→TISSUE DAMAGE  (2)wherein hv indicates applied electromagnetic radiation and(PHOTOSENSITIZER)* indicates photoactivated photosensitizer. The Type 2mechanism proceeds via a three-step process involving activation of thephotosensitizer by absorption of electromagnetic radiation followed byenergy transfer from the activated photosensitizer to oxygen moleculesin the environment of the target tissue. This energy transfer processgenerates excited state oxygen (¹O₂) which subsequently interacts eitherdirectly or indirectly through Reactive Oxygen Species (ROS) with thetarget tissue so as to cause tissue damage. The Type 2 mechanism can berepresented by the following sequence of reactions:Step 1: PHOTOSENSITIZER+hv→(PHOTOSENSITIZER)*  (3)Step 2: (PHOTOSENSITIZER)*+³O₂(Triplet Oxygen)→¹O₂(Singlet Oxygen)  (4)Step 3: ¹O₂(Singlet Oxygen)+TISSUE→TISSUE DAMAGE  (5)wherein hv indicates applied electromagnetic radiation,(PHOTOSENSITIZER)* indicates photoactivated photosensitizer, ³O₂ isground state triplet oxygen, and ¹O₂ is excited state singlet oxygen. Asshown by reactions 1 and 2, Type I photosensitizers do not require thepresence of oxygen for causing tissue damage, and therefore, areexpected to be more effective than Type II photosensitizers underextremely hypoxic environments often found in solid tumors.

The biological basis of tissue injury brought about by tumorphototherapy agents has been the subject of intensive study. Variousbiochemical mechanisms for tissue damage have been postulated, whichinclude the following: a) cancer cells up-regulate the expression of lowdensity lipoprotein (LDL) receptors, and phototherapy (PDT) agents bindto LDL and albumin selectively; (b) porphyrin-like substances areselectively taken up by proliferative neovasculature; (c) tumors oftencontain increased number of lipid bodies and are thus able to bind tohydrophobic photosensitizers; (d) a combination of “leaky” tumorvasculature and reduced lymphatic drainage causes porphyrinaccumulation; (e) tumor cells may have increased capabilities forphagocytosis or pinocytosis of porphyrin aggregates; (f) tumorassociated macrophages may be largely responsible for the concentrationof photosensitizers in tumors; and (g) cancer cells may undergoapoptosis induced by photosensitizers. Among these mechanisms, (f) and(g) are the most general and, of these two alternatives, there is ageneral consensus that (f) is the most likely mechanism by which thephototherapeutic effect of porphyrin-like compounds is induced.

Not withstanding the numerous benefits of phototherapy, these techniquesare not without some drawbacks. For example, local hypoxia is aninherent consequence of phototherapy under some conditions. Localhypoxia may arise directly from oxygen consumption during treatmentand/or indirectly from disruption of tumor vasculature as a result oftreatment. Tissue hypoxia induces a range of molecular and physiologicalresponses including an angiogenesis response associated with geneactivation. For example, hypoxia mediated gene activation is believed toproceed via stabilization of the transcription factor hypoxia-induciblefactor-1α (HIF-1α), which binds to the HIF-1α response element (HRE) inthe promoter of a number of genes including the vascular endothelialgrowth factor (VEGF) gene. Vascular endothelial growth factor is anangiogenic molecule involved with the induction and maintenance ofneovasculature in solid tumors. Animal studies have documented anincrease in VEGF production and accelerated angiogenesis response afterphototherapeutic treatment. (Momma, T; Hamblin, M. R.; Wu, H. C.; Hasan,T; “Photodynamic Therapy of Orthotropic Prostate Cancer withBenzoporphyrin Derivative: Local Control and Distant Metastasis”, CancerResearch, 58, 5425-5431, December 1998.) As a result of increasedsecretion and stabilization of vascular endothelial growth factor (VEGF)in response to some phototherapy procedures, unwanted tumorigenesis andmetastasis processes can be initiated. Accordingly, a number ofcombination therapy strategies for inhibiting this endogenous angiogenicresponse are being pursued as means to enhance the therapeutic efficacyof phototherapy.

A number of anti-VEGF compounds have been evaluated in the context of acombination therapy for the treatment of age related maculardegeneration. U.S. Patent Publication US 2003/0171320, by D. R. Gruyerand published Sep. 11, 2003, discloses methods for treating ocularneovascular disease using anti-VEGF compounds, including aptamers andantibodies, alone or in combination with photodynamic therapy or thermallaser photocoagulation. This reference discloses a number of anti-VEGFaptamers such as nucleic acid ligands having 2′-F-modified nucleotides,2′-O-methyl nucleotides, and pegylated aptamers and generally refers totherapeutic procedures using VEGF antibodies and fragments thereof. Theclinical results provided suggest that combining administration of ananti-VEGF agent with phototherapy enhances efficacy for treatment of agerelated macular degeneration in certain patients. U.S. PatentPublication US 2003/0026945, by Gomer et al. and published Mar. 7, 2002,discloses methods for photodynamic therapy including administration ofan anti-VEGF agent to improve tumoricidal activity. This referencediscloses a single chain polypeptide, EMAP-II, having anti-angiogenicactivity reportedly capable of inhibiting tumor growth and a dipeptideof L-glutamyl-L-tryptophan, IM862, that reportedly inhibits angiogenesisand VEGF production in monocytic lineage cells. PCT InternationalPublication No. WO 2009/117669, published on Sep. 24, 2009, describestreatment with opioid antagonists and mTOR inhibitors in the context ofcellular proliferation and migration.

Several antibody conjugates have recently been developed includingpegaptanib (Macugen®), bevacizumab and ranibizumab for antagonizing VEGFmediated angiogenesis. Bevacizumab is a full-length humanized monoclonalantibody against vascular endothelial growth factor and has beencommercialized as Avastin®. Ranibizumab is a humanized anti-VEGFantibody fragment derived from bevacizumab which inhibits VEGF activityby competitive binding and has been commercialized as Lucentis®. In thecontext of macular degeneration treatment, Avastin® and Lucentis® havebeen shown to stop abnormal vessels from growing and leaking but oftendon't cause permanent closure, and therefore, injections of either drughave to be given repeatedly. Phototherapy on the other hand has beendemonstrated as effective for permanently closing vessels but can resultin vision loss under some conditions. A combination therapy has recentlybeen developed involving phototherapy with half the amount of laser dosefollowed with an injection of Lucentis® or of Avastin® and optionallywith administration an anti-inflammatory steroid. (Augustin, A J, Puls,S, Offerman I, “Triple Therapy for Choroidal Neovascularization due toAge-Related Macular Degeneration. Vertportfrin PDT, Bevacizumab, andDexamethasone. RETINA 27:133-140, 2007).

One problem with Type 2 phototherapy procedure is the induction ofinflammatory response due to reactive oxygen species produced byphotoexcitation of oxygen by Type 2 photosensitizers. This inflammatoryresponse causes the blood vessels to become more porous and, hence,allows cancer cells to metastasize to other regions. As will begenerally recognized from the foregoing, a need currently exists forenhancing the therapeutic efficacy of phototherapy by preventingmetastasis, and for the treatment of macular degeneration using agentsthat are capable of inhibiting inflammatory response, including, but notlimited to the expression and/or activity of vascular endothelial growthfactor.

SUMMARY

In an aspect, the invention relates generally to combination therapyinvolving Type 1 and/or Type 2 photosensitizers and anti-angiogenesisagents. In an embodiment, the invention provides a class of opioidcompounds, structurally related opioid derivatives and their isomersexhibiting inhibitory activity for VEGF expression for use inphototherapy. Opioid compounds and structurally related opioidderivatives of the invention may be administered in combination withadministration and activation of a phototherapy agent. Embodiments ofthe invention include compositions and related phototherapy methods forscavenging excess reactive oxygen species (ROS) generated in aphototherapy procedure. This aspect of the invention is useful forsuppressing (or preventing) tumor metastasis induced by reactive oxygenspecies (ROS) produced in a phototherapy procedure, for example uponactivation of a Type 1 or Type 2 photosensitizer.

Therapeutic agents of the invention include mixtures of stereoisomersand substantially purified stereoisomers of opioids and structurallyrelated derivatives for increasing therapeutic efficacy of aphototherapy procedure by inhibiting an inflammatory response to Type 1or Type 2 phototherapy procedure and, therefore, may be characterized asanti-inflammatory agents. Certain opioid compounds and structurallyrelated derivatives of the present invention are also useful incombination with phototherapy or as a monotherapy for treating a rangeof cancers and ocular neovascular diseases such as age related maculardegeneration. In an embodiment, the invention includes morphinancompounds, morphinanium n-oxide compounds and morphinanium quarternarycompounds and salts thereof for use in a phototherapy procedure and/orfor the treatment of an ocular neovascular disease or cancer, includingpurified stereoisomers of these compounds such as purified (+)enantiomers and (−) enantiomers.

In an aspect, the invention provides therapeutic agents for use in anphototherapy procedure comprising opioids and structurally relatedopioid derivatives. In an embodiment, for example, the inventionprovides a compound for use in a phototherapy procedure, the compoundhaving the formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═O)—, —(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶)—; each of R¹ and R²is independently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl,C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; each of R³-R¹⁶ and R⁵⁰-R⁵⁴ is independently—H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₂-C₈alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀ hydroxyalkyl, orC₅-C₁₀ carbonylalkenylheteroaryl; and n is 0 or 1, wherein: when n is 1,the nitrogen to which R² is attached has a positive charge andoptionally is associated with an anion; and when n is 0, R² is notpresent. In an embodiment, the invention provides N-oxide compoundsuseful as anti-VEGF therapeutic agents for phototherapeutic methodshaving formula (FX1) or (FX2), wherein R² is —(O⁻), and optionally R¹ isC₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₅-C₁₀ aryl or C₂-C₁₀ alkenyl. In anembodiment, for example, the invention provides compounds for use in aphototherapy procedure having any of formula (FX1)-(FX2), wherein eachof R¹ and R² is a group other than H. In an embodiment, for example, theinvention provides compounds for use in a phototherapy procedure havingany of formula (FX1)-(FX2), wherein when n is equal to 0 then R¹ is agroup other than H.

In an aspect, the invention provides therapeutic agents for use in thein viva treatment of an ocular neovascular disease comprising opioids,structurally related opioid derivatives, isomers, N-oxides and saltsthereof. In an embodiment, for example, the invention provides acompound for use in treatment of an ocular neovascular disease havingthe formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶)—; each of R¹ and R² isindependently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; each of R³-R¹⁶ and R⁵⁰-R⁵⁴ is independently—H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₂-C₈alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀ hydroxyalkyl, orC₅-C₁₀ carbonylalkenylheteroaryl; and n is 0 or 1, wherein: when n is 1,the nitrogen to which R² is attached has a positive charge andoptionally is associated with an anion; when n is 0, R² is not present;and when in (FX1) each of X and Y is —OH, n is 1, and W is —(C═O)—, theneach of R¹ and R² is independently a group other than cyclopropylmethyl,and each of R¹ and R² is independently a group other than methyl. In anembodiment, the invention provides N-oxide compounds useful as anti-VEGFtherapeutic agents for phototherapeutic methods having formula (FX1) or(FX2) wherein R² is —(O⁻), and optionally R¹ is C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, C₅-C₁₀ aryl or C₂-C₁₀ alkenyl. In an embodiment, forexample, the invention provides compounds for use in treatment of anocular neovascular disease having any of formula (FX1)-(FX2), whereineach of R¹ and R² is a group other than H. In an embodiment, forexample, the invention provides compounds for use in treatment of anocular neovascular disease having any of formula (FX1)-(FX2), whereinwhen n is equal to 0 then R¹ is a group other than H.

In an aspect, the invention provides therapeutic agents for use in thein vivo treatment of cancer comprising opioids, structurally relatedopioid derivatives, isomers, N-oxides and salts thereof. In anembodiment, for example, the invention provides a compound for thetreatment of cancer, the compound having the formula (FX3) or (FX4):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═O)—, —(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶)—; R¹ is H, —CH₃,—(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; and each of R³-R¹⁶ and R⁵⁰-R⁵⁴ isindependently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈alkenyl, C₂-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl,C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀hydroxyalkyl, or C₅-C₁₀ carbonylalkenylheteroaryl. In an embodiment, theinvention provides N-oxide compounds useful as anti-VEGF therapeuticagents for phototherapeutic methods having formula (FX1) or (FX2)wherein R² is —(O⁻), and optionally R¹ is C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, C₅-C₁₀ aryl, or C₂-C₁₀ alkenyl.

In an embodiment, for example, the invention provides a compound for thetreatment of cancer, the compound having the formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶)—; each of R¹ and R² isindependently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; each of R¹⁰ and R¹¹ is independently —OCH₃,C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₂-C₈ alkoxy, C₅-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, halo,nitrile, C₁-C₈ haloalkyl, C₁-C₁₀ hydroxyalkyl, or C₅-C₁₀carbonylalkenylheteroaryl; each of R³-R⁹, R³-R¹⁶ and R⁵⁰-R⁵⁴ isindependently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈alkenyl, C₂-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl,C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀hydroxyalkyl, or C₅-C₁₀ carbonylalkenylheteroaryl; and n is 0 or 1,wherein: when n is 1, the nitrogen to which R² is attached has apositive charge and optionally is associated with an anion; and when nis 0, R² is not present. In an embodiment, the invention providesN-oxide compounds useful as anti-VEGF therapeutic agents forphototherapeutic methods having formula (FX1) or (FX2) wherein R² is—(O⁻), and optionally R¹ is C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₅-C₁₀ arylor C₂-C₁₀ alkenyl. In an embodiment, for example, the invention providescompounds for use in treatment of cancer having any of formula(FX3)-(FX4), wherein each of R¹ and R² is a group other than H. Theinvention provides compounds for use in treatment of cancer having anyof formula (FX3)-(FX4), wherein when n is equal to 0 then R¹ is a groupother than H.

In an embodiment, the compound of the invention reduces an endogenousangiogenic response to the phototherapy procedure, such as an angiogenicresponse induced by tissue hypoxia. In an embodiment, the compound ofthe invention reduces or prevents migration or proliferation ofendothelial cells in response to the phototherapy procedure, optionallyreducing potential for tumorigenesis and metastasis processes. In anembodiment, the compound of the invention increases the therapeuticefficacy of the phototherapy procedure, for example by increasingtumoricidal activity. In an embodiment, the compound of the invention isfor use in a phototherapy procedure comprising co-administration of theinhibitor agent of VEGF expression and a phototherapy agent, such as aType 1 or Type 2 phototherapy agent, to a target tissue, such as a tumoror other lesion. In an embodiment, the compound of the invention is foruse in a phototherapy procedure for the treatment of cancer or for thetreatment of an ocular neovascular disease such as age related maculardegeneration. In another embodiment, the compound of the invention isadministered with and acts synergistically with the humanized VEGFantibody, its active humanized VEGF antibody fragment and/or an activeanti-VEGF aptamer to increase the therapeutic efficacy of thephototherapy procedure. For example, coadministration of the R isomerderived from (−)naltrexone with a mirror isomer derived from the(+)naltrexone salt is synergistic in combination with VEGF antibody(Avastin) in endothelial cells. (Singleton, P. A.; Garcia, J. G. N. andMoss, J. Mol Cancer Ther 2008; 7(6) 1669-1679).

In an embodiment, the compound of the invention is not methylnaltrexone.In an embodiment of this aspect, the compound of the invention is not a(−) R isomer of methylnaltrexone. In an embodiment, the compound of theinvention is not naltrexone methobromide. In an embodiment, the compoundof the invention is not a (−) R isomer of naltrexone methobromide. In anembodiment, the compound of the invention is not the R isomer derivedfrom (−)naltrexone with a mirror isomer derived from the (+)naltrexone.In an embodiment, the compound of the invention is has a formula otherthan formulas (FX72)-(FX75). In an embodiment of this aspect, thecompound of the invention is has a formula other than formula (FX73). Inan embodiment, the compound is for use in treatment of age relatedmacular degeneration. In an embodiment, the compound of this aspect ofthe invention is for use in a direct monotherapy wherein the compound isadministered to a target tissue to reduce or otherwise alleviateunwanted angiogenesis in connection with an ocular neovascular disease.In such embodiments, the compound is administered withoutco-administration of a phototherapy agent. In another embodiment, thecompound of this aspect of the invention is for use in a combinationtherapy wherein the compound is co-administered with a suitablephototherapy agent that undergoes subsequent excitation upon exposure toelectromagnetic radiation.

The compounds of the present invention include, but are not limited to,therapeutic agents comprising a class of tertiary and quarternaryamines, and salts thereof, for phototherapy and/or for treatment ofocular neovascular disease or cancer. The invention provides, forexample, an opioid or structurally related derivative that is a tertiaryor quaternary amine for phototherapy and/or for treatment of ocularneovascular disease or cancer, the compound having the formula (FX5) to(FX8):

wherein W, X, Y, n, R³-R¹⁶ and R⁵⁰-R⁵⁴ are defined as described inconnection with formulae (FX1) and (FX2). In an embodiment, theinvention provides compounds useful as anti-VEGF therapeutic agents forphototherapeutic methods and/or for treatment of ocular neovasculardisease or cancer having any one of formula (FX5)-(FX8), wherein X is—OH or —OCH₃, and wherein Y is —OH. In an embodiment, the inventionprovides compounds useful as anti-VEGF therapeutic agents forphototherapeutic methods and/or for treatment of ocular neovasculardisease or cancer having any one of formula (FX5)-(FX8), wherein: eachof R¹ and R² is independently allyl or

each q is independently an integer selected from 1 to 4; each r isindependently an integer selected from 1 to 5, and optionally wherein R¹or R² is —CH₃.

The invention also provides, for example, an opioid or structurallyrelated derivative that is a tertiary amine for phototherapy and/or fortreatment of ocular neovascular disease or cancer, the compound havingthe formula (FX9) or (FX10):

wherein X, Y, Ring A and R¹ and R³-R⁷, R⁵⁰, R⁵³, and R⁵⁴ are defined asdescribed in connection with formulae (FX1) and (FX2). The inventionprovides, for example, an opioid or structurally related derivative thatis a quarternary amine or salt thereof for phototherapy and/or fortreatment of ocular neovascular disease or cancer, the compound havingthe formula (FX11) or (FX12):

wherein: each A^(−z) is independently an anion; each z is independently1, 2 or 3; each m is independently 0 or 1, wherein when m is 0, A⁻² isnot present; and wherein X, Y, Ring A, R¹-R⁷, R⁵⁰, R⁵³, and R⁵⁴ aredefined as described in connection with formulae (FX1) and (FX2).

The compounds of the present invention include, but are not limited to,therapeutic agents comprising a class of N-oxides, and salts thereof,for phototherapy and/or for treatment of ocular neovascular disease orcancer. The invention also provides, for example, an opioid orstructurally related derivative that is a quarternary amine N-oxide, orsalt thereof, for phototherapy and/or for treatment of ocularneovascular disease or cancer, the compound having the formula (FX3) or(FX4):

wherein X, Y, Ring A and R¹ and R³-R⁷, R⁵⁰, R⁵³, and R⁵⁴ are defined asdescribed in connection with formulae (FX1) and (FX2). In an embodiment,the invention provides N-oxide compounds useful as anti-VEGF therapeuticagents for phototherapeutic methods and/or for treatment of ocularneovascular disease or cancer having formula (FX3) or (FX4), wherein Xis —OH or —OCH₃, and wherein Y is —OH. In an embodiment, the inventionprovides compounds useful as anti-VEGF therapeutic agents forphototherapeutic methods and/or for treatment of ocular neovasculardisease or cancer having formula (FX3) or (FX4), wherein: each of R¹ andR² is independently allyl or

each q is independently an integer selected from 1 to 4; each r isindependently an integer selected from 1 to 5; and optionally wherein R¹or R² is —CH₃.

In an embodiment, for example, the invention provides a compound for usein a phototherapy procedure or for treatment of an ocular neovasculardisease or cancer, the compound having the formula (FX13)-(FX60):

or a pharmaceutically acceptable salt, solvate, or hydrate, clathrate orprodrug thereof; wherein n, R¹ and R² are defined as described inconnection with formulae (FX1) and (FX2). In an embodiment, theinvention provides compounds useful as anti-VEGF therapeutic agents forphototherapeutic methods or treatment of ocular neovascular disease orcancer having any one of formula (FX13)-(FX60), wherein R¹ or R² isallyl or:

each q is independently an integer selected from 1 to 4; each r isindependently an integer selected from 1 to 5, and optionally wherein R¹or R² is —CH₃. In an embodiment, the invention provides N-oxidecompounds useful as anti-VEGF therapeutic agents having formula(FX13)-(FX60) wherein R² is —(O⁻), and optionally R¹ is C₁-C₁₀ alkyl,C₃-C₁₀ cycloalkyl, C₅-C₁₀ aryl or C₂-C₁₀ alkenyl.

The invention of this aspect also provides purified stereoisomers of thecompounds disclosed herein for use in a phototherapeutic procedure orfor treatment of an ocular neovascular disease or cancer. In anembodiment, for example, the invention provides purified stereoisomercompounds for use in a phototherapeutic procedure or for treatment of anocular neovascular disease or cancer, the compounds having any of theformula:

wherein R¹, R² and n are as described in the context of formulae (FX1)and (FX2).

In an embodiment, for example, the invention provides a compoundcomprising a purified stereoisomer for use in a phototherapy procedureor for treatment of an ocular neovascular disease or cancer, thecompound having any of the formula (FX72) to (FX133):

or a pharmaceutically acceptable salt, solvate, hydrate, clathrate orprodrug thereof. In an embodiment, the invention provides a compound fora phototherapy procedure or for treatment of an ocular neovasculardisease or cancer having any of formula (FX72)-(FX133), provided in asubstantially purified state substantially free of certain otherisomers, such as a substantially purified (+) or (−) stereoisomer. Inembodiments of this aspect, for example, the invention provides acompound for a phototherapy procedure or for treatment of an ocularneovascular disease or cancer having any of formula (FX72)-(FX133),provided in a substantially purified state substantially free of certainother stereoisomers, such as substantially free of other enantiomers. Inan embodiment, for example, the invention provides a compound for aphototherapy procedure or for treatment of an ocular neovascular diseaseor cancer having any of formula (FX72)-(FX133), wherein the compound isprovided as a substantially purified (+) enantiomer or a substantiallypurified (−) enantiomer.

In an embodiment, the invention provides a compound for use in aphototherapy procedure or for use in the treatment of an ocularneovascular disease, the compound having the formula (FX61):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is or—OH;each of R¹ and R² is independently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀carbonylalkenylaryl, or C₅-C₁₀ carbonylalkenylheteroaryl; each ofR²⁰-R²⁹ is independently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl,C₂-C₈ alkenyl, C₁-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl,C₅-C₁₀ carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl; and n is 0 or1, wherein: when n is 1, the nitrogen to which R² is attached has apositive charge and optionally is associated with an anion; and when nis 0, R² is not present.

In an embodiment, the invention provides a compound for the treatment ofcancer, the compound having the formula (FX61):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H,or —OH; each of R¹ and R² is independently —H, —CH₃, —(O⁻), C₁-C₁₀alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀carbonylalkenylaryl, or C₅-C₁₀ carbonylalkenylheteroaryl; each ofR²⁰-R²⁹ is independently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl,C₂-C₈ alkenyl, C₁-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl,C₅-C₁₀ carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl; and n is 0 or1, wherein: when n is 1, the nitrogen to which R² is attached has apositive charge and optionally is associated with an anion; and when nis 0, R² is not present.

In an embodiment, the invention provides a compound for the treatment ofcancer, the compound having the formula (FX62):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H,or —OH;R¹ is —H, —CH₃, C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; and each of R²⁰-R²⁹ is independently —H, —OH,—OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₁-C₈ alkoxy,C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₅-C₁₀carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl. In an embodiment, theinvention provides a compound for the treatment of cancer, the compoundhaving the formula (FX63):

In an embodiment, the invention provides a compound for use in aphototherapy procedure or for use in the treatment of an ocularneovascular disease or cancer, the compound having any of formula(FX64), (FX65), (FX66), or (FX67):

wherein X, Y and R²⁰-R²⁹ are as defined in the context of formula(FX61). In an embodiment, the invention provides a compound for use in aphototherapy procedure or for use in the treatment of an ocularneovascular disease or cancer, the compound having any of formula(FX68), (FX69), (FX70), or (FX71):

wherein X, Y and R²⁰-R²⁹ are as defined in the context of formula(FX61).

The invention of this aspect also provides purified stereoisomers of thecompounds disclosed herein for use in a phototherapeutic procedure orfor treatment of an ocular neovascular disease or cancer. In anembodiment, for example, the invention provides purified stereoisomercompounds for use in a phototherapeutic procedure or for treatment of anocular neovascular disease or cancer, the compounds having any of theformula:

or a pharmaceutically acceptable salt, solvate, hydrate, clathrate orprodrug thereof. In an embodiment, the invention provides a compound fora phototherapy procedure or for treatment of an ocular neovasculardisease or cancer having any of formula (FX134)-(FX164), provided in asubstantially purified state substantially free of certain otherisomers, such as a substantially purified (+) or (−) stereoisomer. Inembodiments of this aspect, for example, the invention provides acompound for a phototherapy procedure or for treatment of an ocularneovascular disease or cancer having any of formula (FX134)-(FX164),provided in a substantially purified state substantially free of certainother stereoisomers, such as substantially free of other enantiomers. Inan embodiment, for example, the invention provides a compound for aphototherapy procedure or for treatment of an ocular neovascular diseaseor cancer having any of formula (FX134)-(FX164), wherein the compound isprovided as a substantially purified (+) enantiomer or a substantiallypurified (−) enantiomer.

In an embodiment, the invention provides compounds useful as anti-VEGFtherapeutic agents for phototherapeutic methods having formula(FX1)-(FX164), wherein X is —OH or —OCH₃, and wherein Y is —OH. In anembodiment, the invention provides compounds useful as anti-VEGFtherapeutic agents for phototherapeutic methods having formula(FX1)-(FX164), wherein R¹ or R² is allyl or

wherein each q is independently an integer selected from 1 to 4; each ris independently an integer selected from 1 to 5, and optionally whereinR¹ or R² is —CH₃ or R² is —(O⁻).

The present invention includes therapeutic agents for phototherapycomprising purified stereoisomers (e.g., enantiomers and diastereomers),salts (including quarternary salts), N-oxides and/or ionic forms (e.g.,protonated and deprotonated forms) of the compounds of formula (FX1) to(FX164), and mixtures thereof. Suitable pharmaceutically acceptablesalts for compounds and compositions of the invention include, withoutlimitation, metal salts such as aluminum, calcium, iron, magnesium,manganese and complex salts, and acid salts such as acetic, aspartic,alkylsulfonic, arylsulfonic, axetil, benzenesulfonic, benzoic,bicarbonic, bisulfuric, bitartaric, butyric, calcium edetate, camsylic,carbonic, chlorobenzoic, cilexetil, citric, edetic, edisylic, estolic,esyl, esylic, formic, fumaric, gluceptic, gluconic, glutamic, glycolic,hexamic, hexylreserinoic, hydrobromic, hydrochloric, hydroiodic,hydroxynaphthoic, isethionic, lactic, lactobionic, maleic, malic,malonic, mandelic, methanesulfonic, mucic, muconic, napsylic, nitric,oxalic, p-nitromethanesulfonic, palmoic, pantothenic, phosphoric,monohydrogen phosphoric, dihydrogen phosphoric, phthalic,polygalactouronic, propionic, salicylic, stearic, succinic, sulfamic,sulfanlic, sulfonic, sulfuric, tannic, tartaric, teoclic,toluenesulfonic, polyglutamic, polyaspartic and the like.

In some embodiments, N-oxides compounds of the invention provideprodrugs of the corresponding base forms. For example, a variety ofenzymes serve to reduce the N-oxide to the active morphinan. [See, e.g.,U.S. Pat. No. 4,722,928 and U.S. Pat. No. 4,990,617]. 3-Hydroxymorphinan N-oxides; such as, N-oxides of naloxone, naltrexone,nalbuphine, nalmefene, pentazocine, butorphanol, buprenorphine,oxymorphone, morphine, dihydromorphine, hydromorphone, levorphanol,levallorphan, and etorphine, may exhibit significantly enhanced oralbioavailability of the corresponding reduced forms.

In an embodiment, the invention provides a compound for use in aphototherapy procedure and/or for use in the treatment of maculardegeneration or cancer, the compound comprising the purified (+)enantiomers of the present opioids and/or derivatives or isomersthereof, such as (+) enantiomer morphinan compounds, (+) enantiomermorphinanium n-oxide compounds or (+) enantiomer morphinaniumquarternary compounds and salts thereof. Methods of phototherapy and/ortreatment of an ocular neovascular disease or cancer using a (+)enantiomer of the present opioids and/or derivatives that issubstantially free of other isomers, such as the (−) enantiomers, isbeneficial for some applications as the purified (+) enantiomer form ofthese compounds does not undergo specific binding to the opioidreceptors. Use of purified (+) enantiomer compounds in the invention,therefore, enable therapeutic methods, including treatment providingeffective anti-VEGF and/or anti-angiogenesis activity, with reducedperipheral and central nervous system effects as compared to use ofcorresponding racemic mixtures and (−) enantiomers. In an embodiment,the invention provides compounds for use in a phototherapy procedureand/or for use in the treatment of macular degeneration or cancer, thecompound being the (+) enantiomer of any of the compounds having any offormula (FX1)-(FX164), wherein the compound is in a purified conditionthat is substantially free of other isomers, for example, substantiallyfree of other stereoisomers such as (−) enantiomers.

In an embodiment, for example, the invention provides compounds for usein a phototherapy procedure and/or for use in the treatment of maculardegeneration or cancer that are not nor-opiate compounds or saltsthereof. In an embodiment, for example, the invention provides compoundsfor use in a phototherapy procedure and/or for use in the treatment ofmacular degeneration or cancer having any of formula (FX1)-(FX164),wherein R¹ and R² are each a group other than —H. In an embodiment, forexample, the invention provides compounds for use in a phototherapyprocedure and/or for use in the treatment of macular degeneration orcancer having any of formula (FX1)-(FX164), wherein when n is equal to 0then R¹ is a group other than —H.

Anti-inflammatory agents of the present invention include therapeuticagents exhibiting in vivo anti-angiogenic activity and/or in vivoanti-VEGF activity. In an embodiment, for example, the inventionprovides an opioid or structurally related opioid derivative for use ina phototherapy procedure or in the treatment of an ocular neovasculardisease or cancer that is an anti-angiogenesis agent. In an embodiment,for example, the invention provides an opioid or structurally relatedopioid derivative for use in a phototherapy procedure or in thetreatment of an ocular neovascular disease or cancer that inhibits VEGFactivity, optionally by inhibiting VEGF mediated intracellularsignaling, for example by inhibiting VEGF receptor tyrosinephosphorylation and/or inhibiting VEGF mediated RhoA activation. Theinvention also includes opioids or structurally related opioidderivatives for use in a phototherapy procedure or in the treatment ofan ocular neovascular disease or cancer that inhibit expression of VEGFor attenuates VEGF-induced VEGF receptor activation.

Anti-inflammatory agents of the present invention include a class ofopioids, including opioid antagonists, opioid agonists and structurallyrelated derivatives and isomers thereof, that exhibit anti-VEGF activityand/or anti-angiogenesis activity. In an embodiment, an opioidantagonist of the invention for a phototherapy procedure or treatment ofan ocular neovascular disease or cancer is a quaternary or N-oxidederivative of (−) or (+) naltrexone, epimers of naltrexol, naloxone,nalmefene, or a quarternary derivative or a pharmaceutically acceptablesalt thereof. For example, an opioid agonist of the invention for somephototherapy procedures and treatment of an ocular neovascular diseaseor cancer is methylnaltrexone or a salt of methylnaltrexone, such asnaltrexone methobromide. In an embodiment, an opioid agonist of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a quarternary derivative or N-oxide ofhydromorphone, epimers of naltrexol, nalmefene, nalorphine, nalbuphine,oxymorphone, oxycodone, hydrocodone, levorphanol, or a pharmaceuticallyacceptable salt thereof. In an embodiment, for example, a structurallyrelated morphinan derivative of the invention for a phototherapyprocedure or treatment of an ocular neovascular disease or cancer is aquaternary salt of dextrorphan, dextromethorphan, levorphanol,dextrallorphan, levallorphan, sinomenine, tetrahydrosinomenine, or aderivative or a pharmaceutically acceptable salt thereof.

Anti-inflammatory agents and VEGF inhibitors of the present inventionmay be co-administered with one or more other therapeutic agents. In anembodiment useful for the treatment of cancer, for example, an opioidcompound or structurally related derivative of the present invention isco-administered with a cytotoxic agent. For example, themethylnaltrexone potentiates ˜700× the cytotoxic effect of5-fluorouracil. Combination therapy of cytotoxics with agents describedherein is effective in treating cancers. (Singleton, P. A.; Garcia, J.G. N. and Moss, J. Mol Cancer Ther 2008; 7(6) 1669-1679.)

The invention also provides therapeutic agents for a phototherapyprocedure or treatment of an ocular neovascular disease or cancercomprising a purified stereoisomer of an opioid or a quarternaryderivative or a pharmaceutically acceptable salt thereof that issubstantially free of other stereoisomers. Advantages of therapeuticagents comprising a purified stereoisomer may include increased in vivoanti-angiogenesis activity and increased in viva anti-VEGF activity.Other advantages of therapeutic agents comprising a purifiedstereoisomer may include improved administration and targeted deliveryperformance and/or enhanced pharmacokinetic and toxicitycharacteristics.

In an embodiment of this aspect, for example, a composition of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified naltrexone stereoisomerquarternary derivative or N-oxide, or a pharmaceutically acceptable saltthereof, substantially free of other methylnaltrexone stereoisomers,wherein the purified naltrexone stereoisomer is (−)-R-methylnaltrexone,(+)-R-methylnaltrexone, (−)-S-methylnaltrexone, or(+)-S-methylnaltrexone.

In an embodiment of this aspect, for example, a compound of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified naloxone stereoisomerquarternary derivative or N-oxide, or a pharmaceutically acceptable saltthereof, substantially free of other naloxone stereoisomers; wherein thepurified naloxone stereoisomer is (+)-R-methylnaloxone,(+)-S-methylnaloxone, (−)-S-methylnaloxone, or (−)-R-methylnaloxone.

In an embodiment of this aspect, for example, a compound of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified nalmefene stereoisomerquarternary derivative, or N-oxide, or a pharmaceutically acceptablesalt thereof, substantially free of other nalmefene stereoisomers,wherein the nalmefene stereoisomer is (+)-R-methlylnalmefene,(+)-S-methylnalmefene, (−)-R-methylnalmefene, or (−)-S-methylnalmefene.

In an embodiment of this aspect, for example, a compound of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified nalbuphine stereoisomer, ora-quarternary derivative, or N-oxide thereof, or a pharmaceuticallyacceptable salt thereof substantially free of other nalbuphinestereoisomers, wherein the nalbuphine stereoisomer is(+)-R-methylnalbuphine, (+)-S-methylnalbuphine, (−)-R-methylnalbuphine,or (−)-S-methylnalbuphine.

In an embodiment of this aspect, for example, a compound of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified oxymorphone or a quarternaryderivative thereof, or N-oxide thereof, substantially free of otheroxymorphone stereoisomers, wherein the oxymorphone stereoisomer is (+)R—or S—N-alkyloxymorphone or (−)R— or S—N-alkyloxymorphone.

In an embodiment of this aspect, for example, a compound of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified hydrocodone stereoisomer ora quarternary derivative, or a N-oxide thereof, or a pharmaceuticallyacceptable salt thereof substantially free of other hydrocodonestereoisomers, wherein the hydrocodone stereoisomer is (+)R— orS—N-alkylhydrocodone or (−)R— or S—N-alkylhydrocodone.

In an embodiment of this aspect, for example, a compound of theinvention for a phototherapy procedure or treatment of an ocularneovascular disease or cancer is a purified oxycodone stereoisomer or aquarternary derivative, or a N-oxide thereof, or a pharmaceuticallyacceptable salt thereof substantially free of other oxycodonestereoisomers, wherein the oxycodone stereoisomer is (+)R— orS—N-alkyloxycodone or (−)R or S—N-alkyloxycodone.

In an embodiment, for example, a compound of the invention for aphototherapy procedure or treatment of an ocular neovascular disease orcancer is a purified hydromorphone stereoisomer or a quarternaryderivative, or a N-oxide thereof, or a pharmaceutically acceptable saltthereof substantially free of other hydromorphone stereoisomers, whereinthe hydromorphone stereoisomer is (+)-hydromorphone or(−)-hydromorphone.

In an aspect, the invention provides a composition for use in aphototherapy procedure, wherein the composition comprises a mixture oftwo or more compounds having any of formulae (FX1)-(FX164) includingisomers, N-oxides and salts thereof, and optionally a Type 1 or Type 2phototherapy agent. In an aspect, the invention provides a compositionfor use in treatment of an ocular neovascular disease or cancer, whereinthe composition comprises a mixture of two or more compounds having anyof formulae (FX1)-(FX164) including isomers, N-oxides and salts thereof.In an embodiment, the invention provides a mixture or formulation foruse in a phototherapy procedure or for use in treatment of an ocularneovascular disease or cancer, wherein at least one of dextrorphan,dextromethorphan, dextrallorphan, (+) cyclorphan, (+) butorphanol,tetrahydrosinomenine, or sinomenine is combined with at least onequarternary derivative or N-oxide of dextrorphan, dextromethorphan,dextrallorphan, (+) cyclorphan, (+) butorphanol, tetrahydrosinomenine,sinomenine or a pharmaceutically acceptable salt thereof. In anembodiment, the invention provides a mixture or formulation for use in aphototherapy procedure or for use in treatment of an ocular neovasculardisease or cancer, wherein at least one of dextrorphan,dextromethorphan, dextrallorphan, (+) cyclorphan, (+) butorphanol,tetrahydrosinomenine, or sinomenine is combined with at least onequaternary derivative or N-oxide of either epimer of naltrexone,naloxone, naltrexol, nalbuphine, nalmefene, hydrocodone, or oxycodone.In an embodiment, the invention provides a mixture or formulation foruse in a phototherapy procedure or for use in treatment of an ocularneovascular disease or cancer, wherein other stereoisomers ofnaltrexone, naloxone, naltrexol, nalbuphine, nalmefene, hydrocodone,oxycodone is combined with at least one quaternary derivative or N-oxideof dextrorphan, dextromethorphan, dextrallorphan, (+) cyclorphan, (+)butorphanol, tetrahydrosinomenine, or sinomenine.

The invention also provides a compound or composition described herein,or a pharmaceutical formulation thereof, for use in a medicalphototherapy procedure comprising: administering to a subject in need oftreatment a therapeutically effective amount of the compound orcomposition; administering to a subject in need of treatment atherapeutically effective amount of a phototherapy agent, and exposingthe administered phototherapy agent to electromagnetic radiation. In anaspect, the phototherapy agent is a Type 1 or Type 2 phototherapy agent.The invention provides a compound or composition described herein, or apharmaceutical formulation thereof, for use in a medical procedure fortreatment of an ocular neovascular disease or cancer comprising:administering to a subject in need of treatment a therapeuticallyeffective amount of the compound or composition described herein. In anaspect, the medical phototherapy procedure or medical procedure fortreatment of an ocular neovascular disease or cancer comprisesadministrating to the subject a therapeutically effective amount of oneor more additional therapeutic agents or diagnostic agents, wherein theone or more additional therapeutic agents or diagnostic agents is one ormore alkylating agents, anti-metabolites, anti-cytoskeletal agents,topoisomerase inhibitors, anti-hormonal agents, or targeted therapeuticagents. In an embodiment, the invention provides a method comprisingcontacting a cell expressing VEGF with a compound having any of formula(FX1)-(FX164), for example for a phototherapy procedure, for thetreatment of an ocular neovascular disease (e.g., MD or AMD) or fortreatment of cancer. In an embodiment, the invention provides a methodcomprising contacting a target tissue with a compound having any offormula (FX1)-(FX164), for example contacting a cancer cell or tumorwith a compound having any of formula (FX1)-(FX164).

In an embodiment, the invention provides a method of treatment,comprising administering to a subject with a disorder characterized byunwanted migration or proliferation of endothelial cells an effectiveamount of any compound or composition described herein.

Without wishing to be bound by any particular theory, there can bediscussion herein of beliefs or understandings of underlying principlesor mechanisms relating to the invention. It is recognized thatregardless of the ultimate correctness of any explanation or hypothesis,an embodiment of the invention can nonetheless be operative and useful.Various features discussed herein in relation to one or more of theexemplary embodiments may be incorporated into any of the describedaspects of the present invention alone or in any combination.

Certain exemplary aspects of the invention are set forth herein. Itshould be understood that these aspects are presented merely to providethe reader with a brief summary of certain forms the invention mighttake and that these aspects are not intended to limit the scope of theinvention. Indeed, the invention may encompass a variety of aspects thatmay not be explicitly set forth herein as would be understood by one ofordinary skill in the relevant art without undue experimentation.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B provide schematic illustrations of methods of thepresent invention for a phototherapy procedure involving (1A)administration of a phototherapy agent without an anti-VEGF therapeuticagent and (1B) co-administration of a phototherapy agent and ananti-VEGF therapeutic agent. Some of these compounds inhibit theexpression by VEGF but are not believed to neutralize VEGF directly likean antibody of VEGF.

FIG. 2 provides a flow diagram comparing phototherapy procedures withand without a step of co-administration of an anti-VEGF agent.

FIG. 3 provides a flow diagram diagrams illustrating methods of thepresent invention for the treatment of an ocular neovascular diseasesincluding the step of administering an anti-VEGF agent. As shown in thisfigure the present methods and compositions for treating ocularneovascular disease encompass both monotherapies and combinationtherapies in conjunction with administration and excitation of aphototherapy agent.

FIG. 4 provides local tumor burden experimental results for comparativestudies involving administration of MNTX, Avastin and Gemcitabineanti-tumor agents. Tumor weight (mg) is shown for tumor conditions of:no treatment (NT); treatment with light alone (light alone); treatmentwith Gemcitabine alone (GEM); treatment with Avastin alone (Avastin) andtreatment with MNTX (MNTX).

FIG. 5 provides a schematic illustrating the experimental design of thepancreatic tumor growth inhibition studies.

FIG. 6 provides experimental results for the pancreatic tumor growthinhibition studies showing the effects of MNTX1 on AsPC-1 pancreaticcancer. FIG. 6 shows tumor weight for control conditions correspondingto intraperitoneal injection of PBS and conditions for corresponding tointraperitoneal injection of MNTX1.

FIG. 7 provides AsPC1 pancreatic tumor size photograph records at theend point of the mouse sacrifice observed for pancreatic tumor growthinhibition studies.

STATEMENTS REGARDING CHEMICAL COMPOUNDS AND NOMENCLATURE

In an embodiment, a composition or compound of the invention is isolatedor purified. In an embodiment, an isolated or purified compound can beat least partially isolated or purified as would be understood in theart. In an embodiment, the composition or compound of the invention hasa chemical purity of 95%, optionally for some applications 99%,optionally for some applications 99.9%, optionally for some applications99.99%, and optionally for some applications 99.999% pure.

Many of the molecules disclosed herein contain one or more ionizablegroups. Ionizable groups are groups from which a proton can be removed(e.g., —COOH) or added (e.g., amines) or which can be quaternized (e.g.,amines). All possible ionic forms of such molecules and salts thereofare intended to be included individually in the disclosure herein. Withregard to salts of the compounds herein, one of ordinary skill in theart can select from among a wide variety of available counterions thatare appropriate for preparation of salts of this invention for a givenapplication. In specific applications, the selection of a given anion orcation for preparation of a salt can result in increased or decreasedsolubility of that salt.

The compounds of this invention can contain one or more chiral centers.Accordingly, this invention is intended to include racemic mixtures,diasteromers, enantiomers, tautomers and mixtures enriched in one ormore stereoisomer. The scope of the invention as described and claimedencompasses the racemic forms of the compounds as well as the individualenantiomers and non-racemic mixtures thereof.

As used herein, the term “group” may refer to a functional group of achemical compound. Groups of the present compounds refer to an atom or acollection of atoms that are a part of the compound. Groups of thepresent invention may be attached to other atoms of the compound via oneor more covalent bonds. Groups may also be characterized with respect tovalence state. The present invention includes groups characterized asmonovalent, divalent, trivalent etc. valence states.

As is customary and well known in the art, hydrogen atoms in formulas(FX1)-(FX164) are not always explicitly shown, for example, hydrogenatoms bonded to the carbon atoms of aromatic, heteroaromatic, andalicyclic rings are not always explicitly shown in formulas(FX1)-(FX164). The structures provided herein, for example in thecontext of the description of formulas (FX1)-(FX164), are intended toconvey to one of reasonable skill in the art the chemical composition ofcompounds of the methods and compositions of the invention, and as willbe understood by one of skill in the art, the structures provided do notindicate the specific bond angles between atoms of these compounds.

As used herein, the term “alkylene” refers to a divalent radical derivedfrom an alkyl group as defined herein. Alkylene groups in someembodiments function as attaching and/or spacer groups in the presentcompositions. Compounds of the invention may have substituted andunsubstituted C₁-C₂₀ alkylene, C₁-C₁₀ alkylene and C₅ alkylene groups.

As used herein, the term “cycloalkylene” refers to a divalent radicalderived from a cycloalkyl group as defined herein. Cycloalkyl groups insome embodiments function as attaching and/or spacer groups in thepresent compositions. Compounds of the invention may have substitutedand unsubstituted C₁-C₂₀ cycloalkyl, C₁-C₁₀ cycloalkyl and C₁-C₅cycloalkyl groups.

As used herein, the term “arylene” refers to a divalent radical derivedfrom an aryl group as defined herein. In some embodiments, an arylene isa divalent group derived from an aryl group by removal of hydrogen atomsfrom two intra-ring carbon atoms of an aromatic ring of the aryl group.Arylene groups in some embodiments function as attaching and/or spacergroups in the present compositions. Arylene groups in some embodimentsfunction as chromophore, fluorophore, aromatic antenna, dye and/orimaging groups in the present compositions. Compounds of the inventioninclude substituted and unsubstituted C₁-C₃₀ arylene, C₁-C₂₀ arylene,C₁-C₁₀ arylene and C₁-C₅ arylene groups.

As used herein, the term “heteroarylene” refers to a divalent radicalderived from a heteroaryl group as defined herein. In some embodiments,an heteroarylene is a divalent group derived from an heteroaryl group byremoval of hydrogen atoms from two intra-ring carbon atoms or intra-ringnitrogen atoms of a heteroaromatic or aromatic ring of the heteroarylgroup. Heteroarylene groups in some embodiments function as attachingand/or spacer groups in the present compositions. Heteroarylene groupsin some embodiments function as chromophore, aromatic antenna,fluorophore, dye and/or imaging groups in the present compositions.Compounds of the invention include substituted and unsubstituted C₁-C₃₀heteroarylene, C₁-C₂₀ heteroarylene, C₁-C₁₀ heteroarylene and C₁-C₅heteroarylene groups.

As used herein, the term “cycloalkylene” refers to a divalent radicalderived from a cycloalkyl group as defined herein. Cycloalkylene groupsin some embodiments function as attaching and/or spacer groups in thepresent compositions. Compounds of the invention include substituted andunsubstituted C₁-C₂₀ cycloalkylene, C₁-C₁₀ cycloalkylene and C₁-C₅cycloalkylene groups.

As used herein, the term “alkenylene” refers to a divalent radicalderived from an alkenyl group as defined herein. Alkenylene groups insome embodiments function as attaching and/or spacer groups in thepresent compositions. Compounds of the invention include substituted andunsubstituted C₁-C₂₀ alkenylene, C₁-C₁₀ alkenylene and C₁-C₅ alkenylenegroups.

As used herein, the term “cylcoalkenylene” refers to a divalent radicalderived from a cylcoalkenyl group as defined herein. Cycloalkenylenegroups in some embodiments function as attaching and/or spacer groups inthe present compositions. Compounds of the invention include substitutedand unsubstituted C₁-C₂₀ cylcoalkenylene, C₁-C₁₀ cylcoalkenylene andC₁-C₅ cylcoalkenylene groups.

As used herein, the term “alkynylene” refers to a divalent radicalderived from an alkynyl group as defined herein. Alkynylene groups insome embodiments function as attaching and/or spacer groups in thepresent compositions. Compounds of the invention include substituted andunsubstituted C₁-C₂₀ alkynylene, C₁-C₁₀ alkynylene and C₁-C₅ alkynylenegroups.

As used herein, the term “halo” refers to a halogen group such as afluoro (—F), chloro (—Cl), bromo (—Br), or iodo (—I).

The term “heterocyclic” refers to ring structures containing at leastone other kind of atom, in addition to carbon, in the ring. Examples ofsuch atoms include nitrogen, oxygen and sulfur. Examples of heterocyclicrings include, but are not limited to, pyrrolidinyl, piperidyl,imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl,pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl,imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl,benzothiadiazolyl, triazolyl and tetrazolyl groups.

The term “carbocyclic” refers to ring structures containing only carbonatoms in the ring. Carbon atoms of carbocyclic rings can be bonded to awide range of other atoms and functional groups.

The term “alicyclic” refers to a ring that is not an aromatic ring.Alicyclic rings include both carbocyclic and heterocyclic rings.

As used herein, the term “alkoxyalkyl” refers to a substituent of theformula alkyl-O-alkyl.

As used herein, the term “polyhydroxyalkyl” refers to a substituenthaving from 2 to 12 carbon atoms and from 2 to 5 hydroxyl groups, suchas the 2,3-dihydroxypropyl, 2,3,4-trihydroxybutyl or2,3,4,5-tetrahydroxypentyl residue.

As used herein, the term “polyalkoxyalkyl” refers to a substituent ofthe formula alkyl-(alkoxy)_(n)-alkoxy wherein n is an integer from 1 to10, preferably 1 to 4, and more preferably for some embodiments 1 to 3.

As used herein, the term “allyl” refers to a substituent of the formula—CH₂CH═CH₂.

Amino acids include glycine, alanine, valine, leucine, isoleucine,methionine, proline, phenylalanine, tryptophan, asparagine, glutamine,glycine, serine, threonine, serine, rhreonine, asparagine, glutamine,tyrosine, cysteine, lysine, arginine, histidine, aspartic acid andglutamic acid. As used herein, reference to “a side chain residue of anatural α-amino acid” specifically includes the side chains of theabove-referenced amino acids.

Alkyl groups include straight-chain, branched and cyclic alkyl groups.Alkyl groups include those having from 1 to 30 carbon atoms. The termcycloalkyl refers to an alky group having a ring structure, such as a.Alkyl groups include small alkyl groups having 1 to 3 carbon atoms.Alkyl groups include medium length alkyl groups having from 4-10 carbonatoms. Alkyl groups include long alkyl groups having more than 10 carbonatoms, particularly those having 10-30 carbon atoms. Cyclic alkyl groupsinclude those having one or more rings. Cycloalkyl groups include thosehaving a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-member carbon ring andparticularly those having a 3-, 4-, 5-, 6-, or 7-member ring. The carbonrings in cyclic alkyl groups can also carry alkyl groups. Cycloalkylgroups can include bicyclic and tricyclic alkyl groups. Alkyl groups areoptionally substituted. Substituted alkyl groups include among othersthose which are substituted with aryl groups, which in turn can beoptionally substituted. Specific alkyl groups include methyl, ethyl,n-propyl, iso-propyl, cyclopropyl, n-butyl, s-butyl, t-butyl,cyclobutyl, n-pentyl, branched-pentyl, cyclopentyl, n-hexyl, branchedhexyl, and cyclohexyl groups, all of which are optionally substituted.Substituted alkyl groups include fully halogenated or semihalogenatedalkyl groups, such as alkyl groups having one or more hydrogens replacedwith one or more fluorine atoms, chlorine atoms, bromine atoms and/oriodine atoms. Substituted alkyl groups include fully fluorinated orsemifluorinated alkyl groups, such as alkyl groups having one or morehydrogen atoms replaced with one or more fluorine atoms. An alkoxy groupis an alkyl group that has been modified by linkage to oxygen and can berepresented by the formula R—O and can also be referred to as an alkylether group. Examples of alkoxy groups include, but are not limited to,methoxy, ethoxy, propoxy, butoxy and heptoxy. Alkoxy groups includesubstituted alkoxy groups wherein the alky portion of the groups issubstituted as provided herein in connection with the description ofalkyl groups. As used herein MeO— refers to CH₃O—.

Alkenyl groups include straight-chain, branched and cyclic alkenylgroups. Alkenyl groups include those having 1, 2 or more double bondsand those in which two or more of the double bonds are conjugated doublebonds. Alkenyl groups include those having from 2 to 20 carbon atoms.Alkenyl groups include small alkenyl groups having 2 to 3 carbon atoms.Alkenyl groups include medium length alkenyl groups having from 4-10carbon atoms. Alkenyl groups include long alkenyl groups having morethan 10 carbon atoms, particularly those having 10-20 carbon atoms.Cyclic alkenyl groups include those having one or more rings. Cyclicalkenyl groups include those in which a double bond is in the ring or inan alkenyl group attached to a ring. Cyclic alkenyl groups include thosehaving a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-member carbon ring andparticularly those having a 3-, 4-, 5-, 6- or 7-member ring. The carbonrings in cyclic alkenyl groups can also carry alkyl groups. Cyclicalkenyl groups can include bicyclic and tricyclic alkyl groups. Alkenylgroups are optionally substituted. Substituted alkenyl groups includeamong others those which are substituted with alkyl or aryl groups,which groups in turn can be optionally substituted. Specific alkenylgroups include ethenyl, prop-1-enyl, prop-2-enyl, cycloprop-1-enyl,but-1-enyl, but-2-enyl, cyclobut-1-enyl, cyclobut-2-enyl, pent-1-enyl,pent-2-enyl, branched pentenyl, cyclopent-1-enyl, hex-1-enyl, branchedhexenyl, cyclohexenyl, all of which are optionally substituted.Substituted alkenyl groups include fully halogenated or semihalogenatedalkenyl groups, such as alkenyl groups having one or more hydrogensreplaced with one or more fluorine atoms, chlorine atoms, bromine atomsand/or iodine atoms. Substituted alkenyl groups include fullyfluorinated or semifluorinated alkenyl groups, such as alkenyl groupshaving one or more hydrogen atoms replaced with one or more fluorineatoms.

Aryl groups include groups having one or more 5-, 6- or 7-memberaromatic and/or heterocyclic aromatic rings. The term heteroarylspecifically refers to aryl groups having at least one 5-, 6- or7-member heterocyclic aromatic rings. Aryl groups can contain one ormore fused aromatic and heteroaromatic rings or a combination of one ormore aromatic or heteroaromatic rings and one or more nonaromatic ringsthat may be fused or linked via covalent bonds. Heterocyclic aromaticrings can include one or more N, O, or S atoms in the ring. Heterocyclicaromatic rings can include those with one, two or three N atoms, thosewith one or two O atoms, and those with one or two S atoms, orcombinations of one or two or three N, O or S atoms. Aryl groups areoptionally substituted. Substituted aryl groups include among othersthose which are substituted with alkyl or alkenyl groups, which groupsin turn can be optionally substituted. Specific aryl groups includephenyl, biphenyl groups, pyridyl, quinolyl, isoquinolyl, pyridazinyl,pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl,benzoxadiazolyl, benzothiadiazolyl, and naphthyl groups, all of whichare optionally substituted. Substituted aryl groups include fullyhalogenated or semihalogenated aryl groups, such as aryl groups havingone or more hydrogens replaced with one or more fluorine atoms, chlorineatoms, bromine atoms and/or iodine atoms. Substituted aryl groupsinclude fully fluorinated or semifluorinated aryl groups, such as arylgroups having one or more hydrogens replaced with one or more fluorineatoms. Aryl groups include, but are not limited to, aromaticgroup-containing or heterocylic aromatic group-containing groupscorresponding to any one of the following: benzene, naphthalene,naphthoquinone, diphenylmethane, fluorene, anthracene, anthraquinone,phenanthrene, tetracene, tetracenedione, pyridine, quinoline,isoquinoline, indoles, isoindole, oxazole, thiazole, pyrazine,benzimidazole, benzofuran, dibenzofuran, carbazole, acridine, acridone,phenanthridine, thiophene, benzothiophene, dibenzothiophene, xanthene,xanthone, flavone, coumarin, azulene or anthracycline. As used herein, agroup corresponding to the groups listed above expressly includes anaromatic or heterocyclic aromatic radical, including monovalent,divalent and polyvalent radicals, of the aromatic and heterocyclicaromatic groups listed herein are provided in a covalently bondedconfiguration in the compounds of the invention at any suitable point ofattachment. In embodiments, aryl groups contain between 5 and 30 carbonatoms. In embodiments, aryl groups contain one aromatic orheteroaromatic six-membered ring and one or more additional five- orsix-membered aromatic or heteroaromatic ring. In embodiments, arylgroups contain between five and eighteen carbon atoms in the rings. Arylgroups optionally have one or more aromatic rings or heterocyclicaromatic rings having one or more electron donating groups, electronwithdrawing groups and/or targeting ligands provided as substituents.

Arylalkyl groups are alkyl groups substituted with one or more arylgroups wherein the alkyl groups optionally carry additional substituentsand the aryl groups are optionally substituted. Specific alkylarylgroups are phenyl-substituted alkyl groups, e.g., phenylmethyl groups.Alkylaryl groups are alternatively described as aryl groups substitutedwith one or more alkyl groups wherein the alkyl groups optionally carryadditional substituents and the aryl groups are optionally substituted.Specific alkylaryl groups are alkyl-substituted phenyl groups such asmethylphenyl. Substituted arylalkyl groups include fully halogenated orsemihalogenated arylalkyl groups, such as arylalkyl groups having one ormore alkyl and/or aryl groups having one or more hydrogens replaced withone or more fluorine atoms, chlorine atoms, bromine atoms and/or iodineatoms.

As to any of the groups described herein which contain one or moresubstituents, it is understood that such groups do not contain anysubstitution or substitution patterns which are sterically impracticaland/or synthetically non-feasible. In addition, the compounds of thisinvention include all stereochemical isomers arising from thesubstitution of these compounds. Optional substitution of alkyl groupsincludes substitution with one or more alkenyl groups, aryl groups orboth, wherein the alkenyl groups or aryl groups are optionallysubstituted. Optional substitution of alkenyl groups includessubstitution with one or more alkyl groups, aryl groups, or both,wherein the alkyl groups or aryl groups are optionally substituted.Optional substitution of aryl groups includes substitution of the arylring with one or more alkyl groups, alkenyl groups, or both, wherein thealkyl groups or alkenyl groups are optionally substituted.

Optional substituents for any alkyl, alkenyl and aryl group includessubstitution with one or more of the following substituents, amongothers:

-   -   halogen, including fluorine, chlorine, bromine or iodine;    -   pseudohalides, including —CN;    -   —COOR where R is a hydrogen or an alkyl group or an aryl group        and more specifically where R is a methyl, ethyl, propyl, butyl,        or phenyl group all of which groups are optionally substituted;    -   —COR where R is a hydrogen or an alkyl group or an aryl group        and more specifically where R is a methyl, ethyl, propyl, butyl,        or phenyl group all of which groups are optionally substituted;    -   —CON(R)₂ where each R, independently of each other R, is a        hydrogen or an alkyl group or an aryl group and more        specifically where R is a methyl, ethyl, propyl, butyl, or        phenyl group all of which groups are optionally substituted; and        where R and R can form a ring which can contain one or more        double bonds and can contain one or more additional carbon        atoms;    -   —OCON(R)₂ where each R, independently of each other R, is a        hydrogen or an alkyl group or an aryl group and more        specifically where R is a methyl, ethyl, propyl, butyl, or        phenyl group all of which groups are optionally substituted; and        where R and R can form a ring which can contain one or more        double bonds and can contain one or more additional carbon        atoms;    -   —N(R)₂ where each R, independently of each other R, is a        hydrogen, or an alkyl group, or an acyl group or an aryl group        and more specifically where R is a methyl, ethyl, propyl, butyl,        phenyl or acetyl group, all of which are optionally substituted;        and where R and R can form a ring which can contain one or more        double bonds and can contain one or more additional carbon        atoms;    -   —SR, where R is hydrogen or an alkyl group or an aryl group and        more specifically where R is hydrogen, methyl, ethyl, propyl,        butyl, or a phenyl group, which are optionally substituted;    -   —SO₂R, or —SOR where R is an alkyl group or an aryl group and        more specifically where R is a methyl, ethyl, propyl, butyl, or        phenyl group, all of which are optionally substituted;    -   —OCOOR where R is an alkyl group or an aryl group;    -   —SO₂N(R)₂ where each R, independently of each other R, is a        hydrogen, or an alkyl group, or an aryl group all of which are        optionally substituted and wherein R and R can form a ring which        can contain one or more double bonds and can contain one or more        additional carbon atoms;    -   —OR where R is H, an alkyl group, an aryl group, or an acyl        group all of which are optionally substituted. In a particular        example R can be an acyl yielding —OCOR″ where R″ is a hydrogen        or an alkyl group or an aryl group and more specifically where        R″ is methyl, ethyl, propyl, butyl, or phenyl groups all of        which groups are optionally substituted.

Specific substituted alkyl groups include haloalkyl groups, particularlytrihalomethyl groups and specifically trifluoromethyl groups. Specificsubstituted aryl groups include mono-, di-, tri, tetra- andpentahalo-substituted phenyl groups; mono-, di-, tri-, tetra-, penta-,hexa-, and hepta-halo-substituted naphthalene groups; 3- or4-halo-substituted phenyl groups, 3- or 4-alkyl-substituted phenylgroups, 3- or 4-alkoxy-substituted phenyl groups, 3- or4-RCO-substituted phenyl, 5- or 6-halo-substituted naphthalene groups.More specifically, substituted aryl groups include acetylphenyl groups,particularly 4-acetylphenyl groups; fluorophenyl groups, particularly3-fluorophenyl and 4-fluorophenyl groups; chlorophenyl groups,particularly 3-chlorophenyl and 4-chlorophenyl groups; methylphenylgroups, particularly 4-methylphenyl groups; and methoxyphenyl groups,particularly 4-methoxyphenyl groups.

DETAILED DESCRIPTION

Referring to the drawings, like numerals indicate like elements and thesame number appearing in more than one drawing refers to the sameelement. In general the terms and phrases used herein have theirart-recognized meaning, which can be found by reference to standardtexts, journal references and contexts known to those skilled in theart. The following definitions are provided to clarify their specificuse in the context of the invention.

“Optical agent” generally refers to compositions, preparations, and/orformulations that absorb, emit, or scatter electromagnetic radiation ofwavelength, generally in the range of 350-1300 nanometers, within abiologically relevant environment or condition. Optical agentsoptionally have molecular recognition or targeting functions enablinglocalized delivery to a target tissue. “Optical imaging agents” or“optical contrast agents” are a class of optical agents that undergoemission via fluorescence or phosphorescence pathways when excited byelectromagnetic radiation. These pathways are useful for diagnosticimaging, visualization, or organ function monitoring. “Phototherapyagents” and “photosensitizers” are used interchangeably and refer to aclass of optical agents that absorb electromagnetic radiation andundergo photochemical reactions, such as photofragmentation of one ormore photolabile bonds, to generate reactive intermediates for achievinga desired therapeutic result. Phototherapy agents include compounds thatabsorbed visible and/or near infrared radiation and generate nitrenes,carbene, free radicals, and/or ions. Phototherapy agents are useful fora wide range of phototherapy applications, for example in the treatmentof tumors or other lesions and in the treatment of age related maculardegeneration. Phototherapy agents include Type 1 and Type 2 phototherapyagents.

“Phototherapy procedure” refers to a therapeutic procedure involvingadministration of a photosensitizer to a patient followed by subsequentexcitation of the phototherapy agent by exposure to appliedelectromagnetic radiation, such as electromagnetic radiation havingwavelengths in the visible and/or near IR region of the electromagneticspectrum, such as wavelengths in the range of 350-1300 nanometers.Phototherapy includes, but is not limited to, photodynamic therapy. Asused herein phototherapy includes procedures involving administration ofType 1 and/or Type 2 phototherapy agents, optionally further includingadministration of one or more additional therapeutic agents, such asanti-angiogenesis agents or anti-VEGF agents.

“Opioid” refers to compounds which bind to opioid receptors. Opioidincludes both opioid agonists and opioid antagonists. Opioid agonistsbind to opioid receptors and alter the function of the receptor, forexample by stimulating the receptor. Opioid agonist include syntheticopioid agonists and semisynthetic opioid agonists. Opioid antagonistsbind to opioid receptors and do not substantially alter the function ofthe receptor, for example by not stimulating the receptor. Opioids ofthe present invention include mu opioid antagonists, mu opioid agonists,kappa opioid antagonists, and kappa opioid agonists. In someembodiments, compositions and methods of the present invention includeperipheral opioids that do not tend to cross the blood brain barrier.Peripheral opioids act primarily on the physiological systems andcomponents external to the central nervous system. In some embodiments,compositions and methods of the present invention include central-actingopioids that readily cross the blood brain barrier central and hence acton physiological systems and components of the central nervous system.

“Angiogenesis” refers molecular and physiological processes involved inthe production of new blood vessels. The formation of new blood cellsvia angiogenesis may involve migration, proliferation anddifferentiation of endothelial cells. “Angiogenic agent” refers to acompound which induces or promotes angiogenesis. “Anti-angiogenic agent”refers to the capability of a compound, or pharmaceutical formulationthereof, to attenuate, inhibits or otherwise reduces angiogenesis,including in some conditions attenuation of the migration, proliferationand differentiation of endothelial cells. Anti-angiogenic agents of thepresent invention may reduce or terminate formation of new blood vesselsin a target tissue. “Proangiogenesis effect” refers to the action of amedical procedure, such as a phototherapy procedure, which acts to causean increase in angiogenesis.

“Target tissue” refers to tissue of a subject to which an opioidcompound, structurally related opioid derivative or isomer thereofand/or phototherapy agent is administered or otherwise contacted, forexample during a medical procedure such as a phototherapy procedure,treatment of an ocular neovascular disease, and/or treatment of cancer.Target tissue may be contacted with an opioid compound, structurallyrelated opioid derivative or isomer thereof and/or phototherapy agent ofthe invention under in viva conditions or ex vivo conditions. Targettissues in some methods of the invention include cancerous tissue,precancerous tissue, a tumor, a lesion, a site of inflammation, orvasculature tissue. Target tissue in some methods of the inventioninclude a melanoma cell, a breast lesion, a prostate lesion, a lungcancer cell, a colorectal cancer cell, an atherosclerotic plaque, abrain lesion, a blood vessel lesion, a lung lesion, a heart lesion, athroat lesion, an ear lesion, a rectal lesion, a bladder lesion, astomach lesion, an intestinal lesion, an esophagus lesion, a liverlesion, or a pancreatic lesion. Target tissue in some embodiments refersto a selected organ of the subject or component thereof, such as lung,heart, brain, stomach, liver, kidneys, gallbladder, pancreas,intestines, rectum, skin, prostate, ovaries, breast, bladder, bloodvessel, throat, ear, or esophagus.

An anti-VEGF agent is a compound which attenuates, inhibits or otherwisereduces VEGF activity in a target tissue. An anti-VEGF agent may inhibitVEGF activity by affecting a range of biological agents and biologicalprocesses. In some embodiments, for example, an anti-VEGF agent inhibitsexpression of VEGF, or directly binds to VEGF in a manner reducing itsability to interact with biological agents in one or more biologicalpathways. In some embodiments, for example, an anti-VEGF agentinterferes or otherwise disrupts VEGF ligand receptor binding, and/orinterferes with post-receptor binding VEGF mediated cellular signaling.In an embodiment, for example, anti-VEGF agents of the invention areopioids and structurally related derivatives that attenuate VEGFactivity by reducing or otherwise interfering with receptor tyrosinephosphorylation and/or VEGF mediated RhoA activation and other pathways.

As used herein, the term “substantially free of other isomers” refers toa composition or pharmaceutical formulation thereof comprising a highmolar ratio of one isomer of a given molecule with respect to some orall other isomers of the given molecule. In some aspects of the presentinvention, for example, a purified isomer substantially free of otherisomers refers to a composition characterized by a molar ratio of thepurified isomer to some or all other isomers present in the compositiongreater than or equal 10, preferably for some embodiments greater thanor equal to 20, preferably for some embodiments greater than or equal to100, preferably for some embodiments greater than or equal to 1000, andpreferably for some embodiments greater than or equal to 10000. Inrelated aspects, the term “substantially free of other isomers” refersto a composition or pharmaceutical formulation thereof comprising a highmolar ratio of one stereoisomer of a given molecule with respect toother stereoisomers of the given molecule. In further related aspects,the term “substantially free of other isomers” refers to a compositionor pharmaceutical formulation thereof comprising a high molar ratio ofone enantiomer of a given molecule with respect to other enantiomers ofthe given molecule. In an embodiment, a substantially purified (+)enantiomer has a molar ratio of the purified (+) enantiomer relative tothe corresponding (−) enantiomer of greater than or equal to 10,preferably for some embodiments greater than or equal to 100, preferablyfor some embodiment greater than or equal to 1000, and preferably forsome embodiments greater than or equal to 10000. In an embodiment, asubstantially purified (−) enantiomer has a molar ratio of the purified(−) enantiomer relative to the corresponding (+) enantiomer of greaterthan or equal to 10, preferably for some embodiments greater than orequal to 100, preferably for some embodiment greater than or equal to1000, and preferably for some embodiments greater than or equal to10000.

“Administering” means that a compound or formulation thereof of thepresent invention, such as an opioid or structurally related opioidderivative, is provided to a patient or subject, for example in atherapeutically effective amount. The present invention includes methodsfor a medical phototherapy procedure comprising administering atherapeutically effective amount of a compound having any one offormulae (FX1)-(FX164) to a patient in need of treatment, for example toa patient undergoing phototherapy or treatment for a diagnosed diseasedstate including cancer or an ocular neovascular disease such as agerelated macular degeneration. “Co-administration” refers toadministering two or more compounds at some time during a biomedicalprocedure. Co-administration refers to administration of two or morecompounds at the same time, or before or after each other during thesame biomedical procedure. Co-administration includes phototherapyprocedures wherein an anti-angiogenesis agent and/or anti-VEGF agent isadministered prior to excitation of the phototherapy agent, duringexcitation of the phototherapy agent and/or after excitation of thephototherapy agent. Co-administration includes phototherapy procedureswherein an anti-angiogenesis agent and/or anti-VEGF agent isadministered multiple times after excitation of the phototherapy agent.

The term “inflammation” generally refers to a biological response ofvascular tissues to harmful stimuli, such as pathogens, damaged cells,irritants, etc. Inflammation can be either acute or chronic. Acuteinflammation is an initial response of the body to harmful stimuli andcan be achieved by the increased movement of plasma and leukocytes fromthe blood into injured tissues. An inflammatory response can involve thelocal vascular system, the immune system, and/or various cells withinthe injured tissue. Prolonged inflammation, referred to as chronicinflammation, can lead to a progressive shift in the type of cells whichare present at the site of inflammation can be characterized bysimultaneous destruction and healing of the tissue from the inflammatoryprocess.

The term “amino acid” comprises naturally occurring amino acids as wellas non-naturally occurring amino acids, including amino acid analogs andderivatives. One skilled in the art will recognize that reference hereinto an amino acid comprises, for example, naturally occurring proteogenicL-amino acids; D-amino acids; chemically modified amino acids such asamino acid analogs and derivatives; naturally occurring non-proteogenicamino acids, and chemically synthesized compounds having propertiesknown in the art to be characteristic of amino acids.

The term “nucleic acid” as used herein generally refers to a molecule orstrand of DNA, RNA, or derivatives or analogs thereof including one ormore nucleobases. Nucleobases comprise purine or pyrimidine basestypically found in DNA or RNA (e.g., adenine, guanine, thymine,cytosine, and/or uracil). The term “nucleic acid” also comprisesoligonucleotides and polynucleotides. Nucleic acids may besingle-stranded molecules, or they may be double-, triple- orquadruple-stranded molecules that may comprise one or more complementarystrands of a particular molecule. “Nucleic acid” includes artificialnucleic acids including peptide nucleic acids, morpholino nucleic acids,glycol nucleic acids and threose nucleic acids. Artificial nucleic acidsmay be capable of nucleic acid hybridization.

As used herein, “sequence” means the linear order in which monomersoccur in a polymer, the order of amino acids in a polypeptide or theorder of nucleotides in a polynucleotide for example.

The terms “peptide” and “polypeptide” are used synonymously in thepresent description, and refer to a class of compounds comprising ofamino acid residues chemically bonded together by amide bonds (orpeptide bonds), regardless of length, functionality, environment, orassociated molecule(s). Peptides and polypeptides are polymericcompounds comprising at least two amino acid residues or modified aminoacid residues. Modifications can be naturally occurring or non-naturallyoccurring, such as modifications generated by chemical synthesis.Modifications to amino acids in peptides include, but are not limitedto, phosphorylation, glycosylation, lipidation, prenylation,sulfonation, hydroxylation, acetylation, methionine oxidation,alkylation, acylation, carbamylation, iodination and the addition ofcofactors. Peptides include proteins and further include compositionsgenerated by degradation of proteins, for example by proteolyicdigestion. Peptides and polypeptides can be generated by substantiallycomplete digestion or by partial digestion of proteins. Polypeptidescomprising 2 to 100 amino acid units, optionally for some embodiments 2to 50 amino acid units and, optionally for some embodiments 2 to 20amino acid units can be used as polypeptide targeting ligands in theinvention, for example, where the polypepetide preferentially binds toproteins, peptides or other biomolecules expressed, or otherwisegenerated by, a target tissue, such as a tumor, precancerous tissue,site of inflammation or other lesion. Typically, the polypeptide is atleast four amino acid residues in length and can range up to afull-length protein.

“Protein” refers to a class of compounds comprising one or morepolypeptide chains and/or modified polypeptide chains. Proteins can bemodified by naturally occurring processes such as post-translationalmodifications or co-translational modifications. Exemplarypost-translational modifications or co-translational modificationsinclude, but are not limited to, phosphorylation, glycosylation,lipidation, prenylation, sulfonation, hydroxylation, acetylation,methionine oxidation, the addition of cofactors, proteolysis, andassembly of proteins into macromolecular complexes. Modification ofproteins can also include non-naturally occurring derivatives, analoguesand functional mimetics generated by chemical synthesis. Exemplaryderivatives include chemical modifications such as alkylation,acylation, carbamylation, halogenation, iodination or any modificationthat derivatizes the protein.

As used herein, “polynucleotide” and “oligonucleotide” are usedinterchangeably and refer to a class of compounds composed of nucleicacid residues chemically bonded together. The invention provides opticalagents having an oligonucleotide or polynucleotide targeting ligandwhich comprises a plurality of nucleic acid residues, such as DNA or RNAresidues, and/or modified nucleic acid residues that preferentiallybinds to proteins, peptides or other biomolecules expressed, orotherwise generated by, a target tissue, such as a tumor, precanceroustissue, site of inflammation or other lesion. Modifications to nucleicacid residues can be naturally occurring or non-naturally occurring,such as modifications generated by chemical synthesis. Oligo- orpoly-nucleotide targeting ligands include, for example, oligo- orpoly-nucleotides comprising 1 to 100 nucleic acid units, optionally forsome embodiments 1 to 50 nucleic acid units and, optionally for someembodiments 1 to 20 nucleic acid units. Polypeptide and oligonucleotideinclude a polymer of at least two nucleotides joined together byphosphodiester bonds and may consist of either ribonucleotides ordeoxyribonucleotides.

The term “aptamer” refers to an oligo- or poly-nucleotide or polypeptidethat binds to, or otherwise selectively or preferentially associateswith, a specific target molecule. For example, the invention providesoptical agents having an aptamer targeting ligand that preferentiallybinds to proteins, peptides or other biomolecules expressed, orotherwise generated by, a target tissue, such as a tumor, precanceroustissue, site of inflammation or other lesion.

“Peptidomimetic” refers to a molecule having activity, includingbiological activity, that resembles that of a polypeptide or issubstantially the same as a polypeptide. Morphine, for example, is apeptidomimetic of endorphin peptide. In some embodiments, apeptidomimetic is a small protein-like polymer designed to mimic thefunctionality of a peptide. Peptidomimetics useful as targeting ligandsfor some compounds of the invention in the present invention includepeptoids and β-peptides. The composition and biological activity ofpeptidomimetics and use of peptidomimetics in targeted diagnostics andtherapeutics are further described in the following references: (1) A.Giannis and T. Kolter, Peptidomimetics for Receptor Ligands—Discovery,Development, and Medical Perspectives, Angewandte Chemie InternationalEdition In English, vol. 32, 1993, pg. 1244-1267 and (2) Patch, J. A. etal., Versatile oligo(N-substituted)glycines: The many roles of peptoidsin drug discovery., Pseudo-Peptides in Drug Discovery 2004, 1-31 and P.E. Nielsen.

“Optical agent” generally refers to compounds, compositions,preparations, and/or formulations that absorb, emit, or scatterelectromagnetic radiation of wavelength generally in the range of350-1300 nanometers, within a biologically relevant environment orcondition. In some embodiments, optical agents of the invention, whenexcited by electromagnetic radiation, undergo emission via fluorescenceor phosphorescence pathways. These pathways are useful for diagnosticimaging, visualization, or organ function monitoring. Compoundsbelonging to this class are commonly referred to as “optical imagingagents” or “optical contrast agents.” In some other embodiments, opticalagents of the invention absorb electromagnetic radiation and undergophotochemical reactions such as photofragmentation of one or morephotolabile bonds to generate reactive species such as nitrenes,carbene, free radicals, ions, excited species, etc. This process isuseful for a wide range of phototherapy applications, for example in thetreatment of tumors or other lesions. Compounds belonging to this classare commonly referred to as “photosensitizers.” The term“photosensitizer” refers to a phototherapy agent or a component thereofproviding for photoactivation, for example, photoactivation resulting ingeneration of reactive species that locally kill, injure, inactivate orotherwise degrade cells (e.g., cancer cells, tumor cells, non-cancercells, etc.). Photosensitizers of some embodiments undergophotoactivation that initiates bond cleavage reactions, such asphotolysis and/or nitrogen extrusion reactions, thereby generatingreactive species capable of causing localized cell death or injury.Optical agents include Type 1 and Type 2 phototherapy agents. Opticalagents include, but are not limited to, phototherapy agents (Type 1 and2), photosensitizers, imaging agents, dyes, detectable agents,photosensitizer agents, photoactivators, and photoreactive agents; andconjugates, complexes, and derivatives thereof.

Optical agents of the present invention can contain fluorophores. Theterm “fluorophore” generally refers to a component or moiety of amolecule or group which causes a molecule or group to be fluorescent.Fluorophores can be functional groups in a molecule which absorbelectromagnetic radiation of first specific wavelengths and re-emitenergy at second specific wavelengths. The amount and wavelengths of theemitted electromagnetic radiation depend on both the fluorophore and thechemical environment of the fluorophore. The term “fluorophore” isabbreviated throughout the present description as “FL”. In aspects ofthe invention, fluorophores emit energy in the visible (e.g. 350 nm to750 nm) and NIR regions (e.g., 750-1300 nm) of the electromagneticspectrum.

As used herein, a “chromophore” is a compound or functional group of acompound that results in absorption of electromagnetic radiation,preferably for some applications electromagnetic radiation havingwavelengths in the UV (e.g. 200 nm to 350 nm) or visible (e.g. 350 nm to750 nm) of the electromagnetic spectrum.

As used herein, a “fluorophore” is a compound or functional group of acompound that results in absorption of electromagnetic radiation andsubsequent fluorescence. Preferably for some applications incorporationof a fluorophore results in compounds of the invention that absorbelectromagnetic radiation and generate fluorescence having wavelengthsin the UV (e.g. 200 nm to 350 nm) or visible (e.g. 350 nm to 750 nm) ofthe electromagnetic spectrum. In some embodiment, incorporation of afluorophore results in compounds having an appreciable quantum yield forfluorescence, such as a quantum yield over the range of 0.001 to 1, 0.01to 1, optionally 0.1 to 1.

As used herein, the term “luminescence” refers to the emission ofelectromagnetic radiation from excited electronic states of atoms ormolecules. Luminescence generally refers to electromagnetic radiationemission, such as photoluminescence, chemiluminescence, andelectrochemiluminescence, among others. In photoluminescence, includingfluorescence and phosphorescence, the excited electronic state iscreated by the absorption of electromagnetic radiation. Luminescencedetection involves detection of one or more properties of theluminescence or associated luminescence process. These properties caninclude intensity, excitation and/or emission spectrum, polarization,lifetime, and energy transfer, among others. These properties can alsoinclude time-independent (steady-state) and/or time-dependent(time-resolved) properties of the luminescence. Representativeluminescence techniques include fluorescence intensity (FLINT),fluorescence polarization (FP), fluorescence resonance energy transfer(FRET), fluorescence lifetime (FLT), total internal reflectionfluorescence (TIRF), fluorescence correlation spectroscopy (FCS),fluorescence recovery after photobleaching (FRAP), and bioluminescenceresonance energy transfer (BRET), among others. By way of example, whenan optical agent is used in the present invention, it is desirable thatthe wavelength of radiation be non-ionizing and be such that it excitesthe optical agent. This excitation can cause a bond of the molecule tobreak and can lead to creation of one or more appropriate radical(s).This excitation can also cause the molecule to emit part of the absorbedenergy at a different wavelength. Such emission can be detected usingfluorometric techniques as described above. One skilled in the art canreadily determine the most appropriate treatment and optional detectiontechnique based, at least in part, on the specific phototherapy agent(s)administered and/or the particular use (e.g., tissue to be treated).

“Optical condition” refers to one or more of the following: thefluorescence quantum yield, fluorescence intensity, fluorescenceexcitation wavelength, wavelength distribution or spectrum, emissionwavelength, wavelength distribution or spectrum, Stokes shift, color,reflectance, phosphorescence, chemiluminescence, scattering, and/orother observable and/or measurable spectral property or phenomenon.

“Phototherapy procedure” refers to a therapeutic procedure involvingadministration of a phototherapy agent to a patient followed bysubsequent excitation by exposure to applied electromagnetic radiation,such as electromagnetic radiation having wavelengths in the visibleand/or near IR region of the electromagnetic spectrum. Such wavelengthscan be in the range of 350-1300 nanometers, so as to generate atherapeutically effective amount of excited phototherapy agent.Phototherapy includes, but is not limited to, photodynamic therapy. Asused herein, “phototherapy” includes procedures involving administrationof Type 1 and/or Type 2 phototherapy agents, optionally furtherincluding administration of one or more additional therapeutic agents.

A detectable optical signal may be, for example, an observable change inabsorbance, reflectance, phosphorescence, chemiluminescence, scattering,or other spectral property.

As used herein, “tumor-specific agent” refers to a compound orcomposition, such as an optical agent, that preferentially accumulatesin a tumor at a higher level than normal tissue regardless of theparticular mechanism of uptake in the tumors, for example, receptormediated or enhanced permeability and retention (EPR). Optical agents ofthe invention include tumor-specific agents, including tumor specificphototherapy agents, for example having a targeting ligand providingspecificity in the administration, delivery and/or binding to tumortissue.

Methods of this invention comprise the step of administering an“effective amount” of the present diagnostic and therapeuticcompositions, formulations and preparations containing the presentcompounds or compositions, to diagnose, image, monitor, evaluate, treat,reduce, alleviate, ameliorate or regulate a biological condition and/ordisease state in a patient. The term “effective amount,” as used herein,refers to the amount of the diagnostic and therapeutic formulation,that, when administered to the individual is effective to diagnose,image, monitor, evaluate, treat, reduce alleviate, ameliorate orregulate a biological condition and/or disease state. As is understoodin the art, an effective amount of a given composition or formulationwill depend at least in part upon the mode of administration (e.g.intravenous, oral, topical administration), any carrier or vehicleemployed, and the specific individual to whom the formulation is to beadministered (age, weight, condition, sex, etc.). The dosagerequirements needed to achieve the “effective amount” vary with theparticular formulations employed, the route of administration, andclinical objectives. Based on the results obtained in standardpharmacological test procedures, projected daily dosages of activecompound or composition can be determined as is understood in the art.

In an embodiment, an effective amount of a compound or composition ofthe invention is a therapeutically effective amount. As used herein, thephrase “therapeutically effective” qualifies the amount of compound orcomposition administered in the therapy. This amount achieves the goalof ameliorating, suppressing, eradicating, preventing, reducing the riskof, or delaying the onset of a targeted condition. In an embodiment, aneffective amount of a compound or composition of the invention is adiagnostically effective amount. As used herein, the phrase“diagnostically effective” qualifies the amount of compound orcomposition administered in diagnosis, for example of a disease state orother pathological condition. The amount achieves the goal of beingdetectable while avoiding adverse side effects found with higher doses.In an embodiment, an active ingredient or other component is included ina therapeutically acceptable amount. In an embodiment, an activeingredient or other component is included in a diagnostically acceptableamount.

It is contemplated that the compounds and pharmaceutically acceptablesalts of the invention can be used as part of a combination. The term“combination” means the administration of two or more compounds directedto a target condition. The treatments of the combination generally canbe co-administered in a simultaneous manner. Two compounds can beco-administered as, for example: (a) a single formulation (e.g., asingle capsule) having a fixed ratio of active ingredients; or (b)multiple, separate formulations (e.g., multiple capsules) for eachcompound. The treatments of the combination can alternatively (oradditionally) be administered at different times.

In certain embodiments, the invention encompasses administering opticalagents useful in the invention to a patient or subject. A “patient” or“subject”, used equivalently herein, refers to an animal. In particular,an animal refers to a mammal, preferably a human. The subject caneither: (1) have a condition able to be monitored, diagnosed, preventedand/or treated by administration of an optical agent of the invention;or (2) is susceptible to a condition that is able to be monitored,diagnosed, prevented and/or treated by administering an optical agent ofthe invention.

When used herein, the terms “diagnosis”, “diagnostic” and other rootword derivatives are as understood in the art and are further intendedto include a general monitoring, characterizing and/or identifying astate of health or disease. The term is meant to encompass the conceptof prognosis. For example, the diagnosis of cancer can include aninitial determination and/or one or more subsequent assessmentsregardless of the outcome of a previous finding. The term does notnecessarily imply a defined level of certainty regarding the predictionof a particular status or outcome.

As defined herein, “administering” means that a compound or formulationthereof of the invention, such as an optical agent, is provided to apatient or subject, for example in a therapeutically effective amount.The invention includes methods for a biomedical procedure wherein atherapeutically or diagnostically effective amount of a compound havingany one of formulas (FX1)-(FX164) is administered to a patient in needof treatment, for example to a patient undergoing treatment for adiagnosed diseased state including cancer and vascular diseases.Administering can be carried out by a range of techniques known in theart including parenteral administration including intravenous,intraperitoneal or subcutaneous injection or infusion, oraladministration, topical or transdermal absorption through the skin, orby inhalation, for example. The chosen route of administration maydepend on such factors as solubility of the compound or composition,location of targeted condition, and other factors which are within theknowledge of one having ordinary skill in the relevant art.

“Topical administration” includes the use of transdermal administration,such as transdermal patches or iontophoresis devices.

“Parenteral administration” includes subcutaneous injections,intravenous injections, intraarterial injections, intraorbitalinjections, intracapsular injections, intraspinal injections,intraperitoneal injections, intramuscular injections, intrasternalinjections, and infusion. Dosage forms suitable for parenteraladministration include solutions, suspensions, dispersions, emulsions,and any other dosage form that can be administered parenterally.

As used herein, the term “controlled-release component” refers to anagent that facilitates the controlled-release of a compound including,but not limited to, polymers, polymer matrices, gels, permeablemembranes, liposomes, microspheres, or the like, or any combinationthereof. Methods for producing compounds in combination withcontrolled-release components are known to those of skill in the art.

As used herein, the term “pharmaceutically acceptable” means approved bya regulatory agency of an appropriate federal or state government; orlisted in the U.S. Pharmacopoeia or other generally recognizedpharmacopoeia for use in animals, and more particularly in humans; ordoes not impart significant deleterious or undesirable effect on asubject to whom it is administered and in the context in which it isadministered.

As will be clear to those of ordinary skill in the art, the groups andstructures described herein as portions of the compounds of theinvention may be defined as if they are separate valence-satisfiedchemical structures. It is intended that when a group is described orshown as being a substituent of another group, that the group be viewedas having a valency to allow this binding to occur.

As to any of the above groups which contain one or more substituents, itis understood, that such groups do not contain any substitution orsubstitution patterns which are sterically impractical and/orsynthetically non-feasible. In addition, the compounds of this inventioninclude all stereochemical isomers arising from the substitution ofthese compounds.

Pharmaceutically acceptable salts comprise pharmaceutically-acceptableanions and/or cations. Pharmaceutically-acceptable cations include amongothers, alkali metal cations (e.g., Li⁺, Na⁺, K⁺), alkaline earth metalcations (e.g., Ca²⁺, Mg²⁺), non-toxic heavy metal cations and ammonium(NH₄ ⁺) and substituted ammonium (N(R′)₄ ⁺, where R′ is hydrogen, alkyl,or substituted alkyl, i.e., including, methyl, ethyl, or hydroxyethyl,specifically, trimethyl ammonium, triethyl ammonium, and triethanolammonium cations). Pharmaceutically-acceptable anions include amongother halides (e.g., Cl⁻, Br⁻), sulfate, acetates (e.g., acetate,trifluoroacetate), ascorbates, aspartates, benzoates, citrates, andlactate. Pharmaceutically acceptable salts include, but are not limitedto, salts derived from quarternary amines, such as quarternary amineopioids and structurally related derivatives.

The compounds of this invention may contain one or more chiral centers.Accordingly, therapeutic agents, including anti-angiogenesis agents andanti-VEGF agents, include racemic mixtures of stereoisomers, andcompositions comprising mixtures enriched in one or more stereoisomerand compositions comprising purified stereoisomers (e.g, diastereomers,enantiomers). The scope of the invention as described and claimedencompasses the racemic forms of the compounds as well as non-racemicmixtures thereof and purified forms of individual stereoisomers of thepresent compounds.

Before the present methods are described, it is understood that thisinvention is not limited to the particular methodology, protocols, celllines, and reagents described, as these may vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular embodiments only, and is not intended to limit thescope of the invention which will be limited only by the appendedclaims.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural reference unless thecontext clearly dictates otherwise. Thus, for example, reference to “acell” includes a plurality of such cells and equivalents thereof knownto those skilled in the art, and so forth. As well, the terms “a” (or“an”), “one or more” and “at least one” can be used interchangeablyherein. It is also to be noted that the terms “comprising”, “including”,and “having” can be used interchangeably.

In certain embodiments, the invention encompasses administeringcompounds, such as anti-angiogenesis agents, anti-VEGF agents andphototherapy agents, useful in the invention to a patient or subject. A“patient” or “subject”, used equivalently herein, refers to an animal.In particular, an animal refers to a mammal, preferably a human. Thesubject may either: (1) have a condition diagnosable, preventable and/ortreatable by administration of an optical agent of the invention; or (2)is susceptible to a condition that is diagnosable, preventable and/ortreatable by administering an optical agent of this invention.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of ordinary skillin the art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. Nothing herein is to be construed as anadmission that the invention is not entitled to antedate such disclosureby virtue of prior invention.

Compositions of the invention includes formulations and preparationscomprising one or more of the present optical agents provided in anaqueous solution, such as a pharmaceutically acceptable formulation orpreparation. Optionally, compositions of the invention further compriseone or more pharmaceutically acceptable surfactants, buffers,electrolytes, salts, carriers, binders, coatings, preservatives and/orexcipients.

In an embodiment, the invention provides a pharmaceutical formulationcomprising a composition of the invention, such as a compound of any oneof formulae (FX1)-(FX164). In an embodiment, the invention provides amethod of synthesizing a composition of the invention or apharmaceutical formulation thereof, such as a compound of any one offormulae (FX1)-(FX164). In an embodiment, a pharmaceutical formulationcomprises one or more excipients, carriers, diluents, and/or othercomponents as would be understood in the art. Preferably, the componentsmeet the standards of the National Formulary (“NF”), United StatesPharmacopoeia (“USP”; United States Pharmacopeial Convention Inc.,Rockville, Md.), or Handbook of Pharmaceutical ManufacturingFormulations (Sarfaraz K. Niazi, all volumes, ISBN: 9780849317521, ISBN10: 0849317525; CRC Press, 2004). See, e.g., United States Pharmacopeiaand National Formulary (USP 30-NF 25), Rockville, Md.: United StatesPharmacopeial Convention; 2007; and 2008, and each of any earliereditions; The Handbook of Pharmaceutical Excipients, published jointlyby the American Pharmacists Association and the Pharmaceutical Press(Pharmaceutical Press (2005) (ISBN-10: 0853696187, ISBN-13:978-0853696186); Merck Index, Merck & Co., Rahway, N.J.; and Gilman etal., (eds) (1996); Goodman and Gilman's: The Pharmacological Bases ofTherapeutics, 8th Ed., Pergamon Press. In embodiments, the formulationbase of the formulations of the invention comprises physiologicallyacceptable excipients, namely, at least one binder and optionally otherphysiologically acceptable excipients. Physiologically acceptableexcipients are those known to be usable in the pharmaceutical technologysectors and adjacent areas, particularly, those listed in relevantpharmacopeias (e.g. DAB, Ph. Eur., BP, NF, USP), as well as otherexcipients whose properties do not impair a physiological use.

In an embodiment, an effective amount of a composition of the inventionis a therapeutically effective amount. In an embodiment, an effectiveamount of a composition of the invention is a diagnostically effectiveamount. In an embodiment, an active ingredient or other component isincluded in a therapeutically acceptable amount. In an embodiment, anactive ingredient or other component is included in a diagnosticallyacceptable amount.

Variations on compositions including salts and ester forms of compounds:Compounds of this invention and compounds useful in the methods of thisinvention include those of the compounds and formula (s) describedherein and pharmaceutically-acceptable salts and esters of thosecompounds. In embodiments, salts include any salts derived from theacids of the formulas herein which acceptable for use in human orveterinary applications. In embodiments, the term esters refers tohydrolyzable esters of compounds of the names and structural formulasherein. In embodiments, salts and esters of the compounds of theformulas herein can include those which have the same or bettertherapeutic, diagnostic, or pharmaceutical (human or veterinary) generalproperties as the compounds of the formulas herein. In an embodiment, acomposition of the invention is a compound or salt or ester thereofsuitable for pharmaceutical formulations.

In an embodiment, the invention provides a method for treating a medicalcondition comprising administering to a subject (e.g. patient) in needthereof, a therapeutically effective amount of a composition of theinvention, such as a compound of any one of formulae (FX1)-(FX164). Inan embodiment, the medical condition is cancer, or various otherdiseases, injuries, and disorders, including cardiovascular disorderssuch as atherosclerosis and vascular restenosis, inflammatory diseases,ophthalmic diseases and dermatological diseases.

In an embodiment, the invention provides a medicament which comprises atherapeutically effective amount of one or more compositions of theinvention, such as a compound of any one of formulas (FX1)-(FX164). Inan embodiment, the invention provides a method for making a medicamentfor treatment of a condition described herein, such as the treatment ofocular neovascular disease, macular degeneration, cancer, inflammation,stenosis or a vascular disease. In an embodiment, the invention providesa method for making a medicament for diagnosis or aiding in thediagnosis of a condition described herein, such as the diagnosis ofocular neovascular disease, macular degeneration, cancer, inflammation,stenosis or a vascular disease. In an embodiment, the invention providesthe use of one or more compositions set forth herein for the making of amedicament for the treatment of ocular neovascular disease, maculardegeneration, cancer, inflammation, stenosis or a vascular disease. Inan embodiment, the invention provides the use of one or morecompositions set forth herein for the treatment of a disease.Compositions of the invention include formulations and preparationscomprising one or more of the present optical agents provided in anaqueous solution, such as a pharmaceutically acceptable formulation orpreparation. Optionally, compositions of the invention further compriseone or more pharmaceutically acceptable surfactants, buffers,electrolytes, salts, carriers, binders, coatings, preservatives and/orexcipients.

Compounds of the invention can have prodrug forms. Prodrugs of thecompounds of the invention are useful in embodiments includingcompositions and methods. Any compound that will be converted in vivo toprovide a biologically, pharmaceutically, diagnostically, ortherapeutically active form of a compound of the invention is a prodrug.Various examples and forms of prodrugs are well known in the art.Examples of prodrugs are found, inter alia, in Design of Prodrugs,edited by H. Bundgaard, (Elsevier, 1985), Methods in Enzymology, Vol.42, at pp. 309-396, edited by K. Widder, et. al. (Academic Press, 1985);A Textbook of Drug Design and Development, edited by Krosgaard-Larsenand H. Bundgaard, Chapter 5, “Design and Application of Prodrugs,” by H.Bundgaard, at pp. 113-191, 1991); H. Bundgaard, Advanced Drug DeliveryReviews, Vol. 8, p. 1-38 (1992); H. Bundgaard, et al., Journal ofPharmaceutical Sciences, Vol. 77, p. 285 (1988); and Nogrady (1985)Medicinal Chemistry A Biochemical Approach, Oxford University Press, NewYork, pages 388-392). A prodrug, such as a pharmaceutically acceptableprodrug can represent prodrugs of the compounds of the invention whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use.Prodrugs of the invention can be rapidly transformed in vivo to a parentcompound of a compound described herein, for example, by hydrolysis inblood or by other cell, tissue, organ, or system processes. Furtherdiscussion is provided in T. Higuchi and V. Stella, Pro-drugs as NovelDelivery Systems, V. 14 of the A.C.S. Symposium Series, and in Edward B.Roche, ed., Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press (1987).

The invention contemplates pharmaceutically active compounds eitherchemically synthesized or formed by in viva biotransformation tocompounds set forth herein.

In an embodiment, a composition of the invention is isolated orpurified. In an embodiment, an isolated or purified compound may be atleast partially isolated or purified as would be understood in the art.

The invention is further detailed in the following Examples, which areoffered by way of illustration and are not intended to limit the scopeof the invention in any manner.

Example 1 Methods of Using Anti-VEGF Opioids and Structurally RelatedDerivatives for Phototherapy

Phototherapy, such as photodynamic therapy (PDT), typically employs acombination of a nontoxic photosensitizer (PS) and visible or nearinfrared light to generate reactive species that kill or otherwisedegrade target cells, such as tumors or other lesions. Many clinicallystudied photosensitizers are Type 2 phototherapy agents based on thetetrapyrrole structure of porphyrins, chlorins, and related moleculesthat generate cytotoxic reactive oxygen species (ROS), e.g. singletoxygen, hydroxyl and hydroperoxyl radicals, and the like. Photodynamictherapy in conjunction with Type 2 phototherapy agents in certain tumoranimal models has been shown under certain conditions to initiatetumorigenesis and metastasis through the up-regulation of vascularendothelial growth factor (VEGF) and increased permeability of damagedvasculature. Localized hypoxia and oxidative stress resulting from thephototherapy process, for example, are believed to initiate anangiogenic response. Since tumorigenesis defines the growth andinvasiveness of tumors, antagonizing this endogenous angiogenic responseto PDT provides a potential means to enhance the efficacy ofphototherapy.

Anti-VEGF antibodies, and fragments thereof, as an inhibitor to VEGFexpression, for example, has been shown to reduce angiogenesis post-PDTwith Type 2 phototherapy agents and to reduce the metastaticprogression. Antibodies are known to elicit anti-idiotypic response,however, that can reduce the effectiveness of immunoagents, therebyincreasing the dose needed for a desired therapeutic outcome. Indeed,anti-idiotypic response is a natural “policing” by the immune system toensure that large amounts of antibodies are not accumulated in the bodythat may lead to autoimmune diseases. Furthermore, because antibodyproduction, isolation, and purification is expensive for the quantitiesthat are needed for many therapeutic applications, there is currently asignificant need for small molecular inhibitors of VEGF expression as anadjunct to phototherapy.

The present invention provides opioids and structurally relatedderivatives useful as anti-angiogenesis agents and anti-VEGF agents formitigating angiogenic and metastatic responses initiated byphototherapy. In some embodiments, for example, the present inventionprovides methods comprising in vivo co-administration of an anti-VEGFagent comprising an opioid or structurally related derivative and a Type1 or Type 2 photosensitizer. FIG. 1A provides a schematic diagramillustrating a phototherapy procedure not including the step ofadministering an anti-VEGF agent. As shown in this figure, aphototherapy agent, such as a Type 2 phototherapy agent, is administeredto a patient. Administration provides delivery of the phototherapy agentto a target tissue, such as a tumor or lesion. The phototherapy agent issubsequently excited by exposure to electromagnetic radiation, forexample electromagnetic radiation having wavelengths selected over therange of 350 nm to 1300 nm. Excitation of the phototherapy agentgenerates reactive species, such as free radicals, that result in celldeath. As indicated in FIG. 1A, an unwanted collateral effect ofadministration and excitation of the phototherapy agent under someconditions is an endogenous angiogenic response, thereby increasing thepotential for metastasis. FIG. 1B provides a schematic diagramillustrating a phototherapy procedure of the present invention includingthe step of administering an anti-VEGF agent. As shown in this figure, aphototherapy agent, such as a Type 2 phototherapy agent, isco-administered to a patient with an anti-VEGF agent, such as an opioid(e.g. MNTX or specific (+) or (−) enantiomer thereof), or a structurallyrelated derivative thereof. Administration provides delivery of thephototherapy agent and anti-VEGF agent to a target tissue, such as atumor or lesion. Similar to the process outlined in FIG. 1A, thephototherapy agent is subsequently excited by exposure toelectromagnetic radiation, thereby generating reactive species resultingin cell death. As shown in FIG. 1B, however, administration of theanti-VEGF agent suppress the endogenous angiogenic response, therebydecreasing the potential for metastasis. FIG. 2 provides a flow diagramcomparing phototherapy procedures with and without a step ofco-administration of an anti-VEGF agent.

The opioid antagonist (−)-R-Naltrexone methobromide (MNTX), is a potentinhibitor of VEGF activity that attenuates vascular permeability due todisease, injury or cancer. Therefore, in specific therapeutic methods ofthe present invention, MNTX serves as an adjunct to phototherapy toeffectively mitigate enhancement and/or the dissemination of cancer. Forexample, the purified enantiomer, (+)-R-Naltrexone methobromide and,optionally purified epimers, (−)-S-Naltrexone methobromide and(+)-S-Naltrexone methobromide, provide anti-VEGF therapeutic agents foradministration in combination with phototherapy. MNTX is described as anexample of an anti-VEGF and anti-angiogenic agent of the invention. Aswill be understood by one of skill in the art, other forms ofnaltrexone, including other naltrexone salts and other naltrexonestereoisomers are useful in the present invention as anti-VEGF andanti-angiogenic agents. The structures of various stereoisomers ofnaltrexone methobromide useful in the invention, for example, areprovided below in Table 1-1. The structures of various stereoisomers ofnaltrexone N-oxide useful in the invention, for example, are providedbelow in Table 1-2.

TABLE 1-1 Stereoisomers of naltrexone methobromide. Item No. NameChemical Structure of Stereoisomers 1 naltrexone methobromide

TABLE 1-2 Isomers of naltrexone N-oxide. Item No. Name ChemicalStructure of N-Oxide isomers 1 naltrexone N-oxide

Anti-VEGF and Anti-angiogenic agents of the methods of this aspect ofthe invention further include other opioid antagonists, opioid agonistsand structurally related derivatives thereof including, but not limitedto, quaternary salts and isomeric and epimeric forms of:(+/−)R/S-Naloxone, (+/−)R/S-Nalmefene, nalbuphine, (+/−)-oxymorphone,(+/−)-hydrocodone, (+/−)-oxycodone, (+/−)-hydromorphone,tetrahydrosinomenine, dextrorphan, and dextromethorphan. These compoundsprovide anti-VEGF therapeutic agents for administration in combinationwith phototherapy. As shown in Table 1-2 the N-oxides may becharacterized as either axial or equatorial.

Structures of additional examples of anti-VEGF and Anti-angiogenicagents are provided in Table 1-3.

TABLE 1-3 Specific Opioids and Related Derivatives Item No. Base NameChemical Structure of Stereoisomers 1 Naloxone

2 Nalorphine

3 14- Deoxynaltrexone

4 Nalmefene

5 Oxymorphone

6 Hydrocodone

7 Oxycodone

8 Hydromorphone

9 Nalbuphine

10 Buprenorphine

11 Sinomenine

12 Tetra- hydrosinomenine

13 Dextrorphan

14 Levorphanol

15 Dextromethorphan

16 Levallorphan

17 Dextrallorphan

18 (−)Cyclorphan

19 (+)Cyclorphan

20 (−)Oxilorphan

21 (+)Oxilorphan

22 (−)Butorphanol

23 (+)Butorphanol

24 Nalfurafine

25 Nalorphine

In the structures provided in Tables 1-1, 1-2, and 1-3, (+) and (−)denote the opposite optical isomers of the opioid compounds orstructurally related derivatives. The R and S isomers created around theprochiral nitrogen atom of the quaternary derivatives have the attachedgroups of the nitrogen (e.g., R¹, R² in formulae (FX1) and (FX2)) eitheraxial or equatorial with respect to the plane of the nitrogen containingcyclohexyl ring. Standard prioritization rules are followed. As will beunderstood by those having skill in the art, the N-alkyl substitution inthe structures above does not have to be restricted to methyl.Therefore, the invention includes compounds having groups other thanmethyl attached to the nitrogen including, but not limited to, —H, —O⁻,C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, or C₅-C₁₀alkylaryl. As will be understood by those having skill in the art, theinvention included N-oxides of the compounds provide in Table 1-2,wherein oxygen is in place of the methyl on the nitrogen. The (+)isomers are believe under some conditions not to bind to the opiatereceptors and, thus, are preferred for some therapeutic applications.The N-oxides are usually axial although in the case of N-methyl amixture is likely that can be separated by chromatography techniquesknow in the art.

The invention further includes compositions comprising mixtures of aplurality of the present opioids and structurally related derivativesexhibiting inhibitory activity for VEGF expression. In an embodiment,for example, a composition of the present invention comprises aplurality of different opioids and/or opioid derivatives for use in aphototherapy procedure or for use in treatment of an ocular neovasculardisease or cancer. In an embodiment, a composition of the presentinvention comprises a plurality of different isomers of a particularopioid or opioid derivative of the invention. In an embodiment, acomposition of the present invention includes a plurality of isomers,N-oxides and or salts of a particular opioid or opioid derivative of theinvention. In an embodiment, a mixture of the invention comprises a baseand an N-oxide or salt of the compounds in Tables 1-1, 1-2, and 1-3.

In an aspect, the invention provides a method for a phototherapyprocedure comprising administration to a patient in need of phototherapya therapeutic agent for scavenging reactive oxygen species (ROS)generated during phototherapy, including Type 2 phototherapy.Therapeutic agents useful in this aspect include anti-inflammatoryagents and anti-angiogenesis agents, such as VEGF inhibitors. In anembodiment, for example, the invention provides a method for aphototherapy procedure, the method comprising: (i) administering to apatient in need of phototherapy an effective amount of a phototherapyagent; (ii) administering to the patient an effective amount of acompound having the formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═O)—, —(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶)—; each of R¹ and R²is independently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl,C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; each of R³-R¹⁵ and R⁵⁰-R⁵⁴ is independently—H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₂-C₈alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀ hydroxyalkyl, orC₅-C₁₀ carbonylalkenylheteroaryl; and n is 0 or 1, wherein: when n is 1,the nitrogen to which R² is attached has a positive charge andoptionally is associated with an anion; and when n is 0, R² is notpresent; and (iii) exposing the phototherapy agent administered to thepatient to electromagnetic radiation.

In an embodiment, the invention provides a method for a phototherapyprocedure, the method comprising: (i) administering to a patient in needof phototherapy an effective amount of a phototherapy agent; (ii)administering to the patient an effective amount of a compound havingthe formula (FX61):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H,or —OH; each of R¹ and R² is independently —H, —CH₃, —(O⁻), C₁-C₁₀alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl,C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀carbonylalkenylaryl, or C₅-C₁₀ carbonylalkenylheteroaryl; each ofR²⁰-R²⁹ is independently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl,C₂-C₈ alkenyl, C₁-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl,C₅-C₁₀ carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl; and n is 0 or1, wherein: when n is 1, the nitrogen to which R² is attached has apositive charge and optionally is associated with an anion; and when nis 0, R² is not present; and (iii) exposing the phototherapy agentadministered to the patient to electromagnetic radiation. In anembodiment of this aspect, the compound having the formula (FX1), (FX2)or (FX61) and the phototherapy agent are administered at the same time(e.g., co-administered). In an embodiment of this aspect, the compoundhaving the formula (FX1), (FX2) or (FX61) is administered before, duringor after administration of the phototherapy agent. In an embodiment ofthis aspect, the compound having the formula (FX1), (FX2) or (FX61) isadministered before, during or after exposure of the phototherapy agentto electromagnetic radiation.

In an embodiment, the method of this aspect of the invention comprisesadministering to a patient an anti-angiogenesis agent and/or anti-VEGFagent. In an embodiment, the invention comprises administering atertiary amine having formula (FX1), (FX2) or (FX61). In an embodiment,the invention comprises administering a quarternary amine or saltthereof having formula (FX1), (FX2) or (FX61). In an embodiment, theinvention comprises administering an N-oxide or salt thereof havingformula (FX1), (FX2) or (FX61). In an embodiment, the method of thisaspect of the invention comprises administering to a patient a compoundhaving any one of formulae (FX1)-(FX164), including all of the specificcomposition classes and compounds described herein, in combination withany of the method steps presented herein. As will be understood by oneof skill in the art, the present methods expressly include administeringone or more anti-VEGF agents, including the compound classes, compounds,and variations thereof described herein, including the compound classes,compounds and variations described in connection with any one offormulae (FX1)-(FX164), in combination with the steps of administeringand exposing the phototherapy agent administered to the patient toelectromagnetic radiation.

In an embodiment, the invention provides a method of alleviating aproangiogenesis effect of a medical therapy, the method comprising:administering to a patient undergoing a phototherapy procedure aneffective amount of a compound having any one of formulae (FX1)-(FX164)or a pharmaceutically acceptable salt, solvate, hydrate, clathrate orprodrug thereof, including all of the specific compositions classes andcompounds described herein. In an embodiment, the invention provides amethod of inhibiting an endogenous angiogenic response to a phototherapyprocedure, the method comprising: administering to a patient undergoinga phototherapy procedure an effective amount of a compound having anyone of formulae (FX1)-(FX164) or a pharmaceutically acceptable salt,solvate, hydrate, clathrate or prodrug thereof. In an embodiment, theinvention provides a method of increasing the therapeutic efficacy of aphototherapy procedure, the method comprising: administering to apatient undergoing the phototherapy procedure an effective amount of acompound having the any one of formulae (FX1)-(FX184) or apharmaceutically acceptable salt, solvate, hydrate, clathrate or prodrugthereof, including all of the specific composition classes and compoundsdescribed herein.

Phototherapy methods of the invention include photodynamic therapy andthermal laser photocoagulation. Methods of the present invention mayfurther comprise a number of additional steps. In an embodiment, forexample, the method further comprises delivering a compound of any oneof formulae (FX1)-(FX164) and phototherapy agent to a target tissue ororgan, such as a tumor. In an embodiment, the method further comprisescontacting a target tissue or organ with a compound of any one offormulae (FX1)-(FX164) and a phototherapy agent. In an embodiment, thepresent methods further comprise the step of administering the compoundof any one of formulae (FX1)-(FX164) and the phototherapy agent into abodily fluid of the subject. Anti-VEGF agents and phototherapy agentsmay be introduced into the patient by any suitable method, includingintravenous, intraperitoneal or subcutaneous injection or infusion, oraladministration, transdermal absorption through the skin, or byinhalation. In an embodiment, the method further comprises allowing theanti-VEGF agent and phototherapy agent to accumulate in a target tissueor organ prior to exposure of the phototherapy agent to electromagneticradiation. In an embodiment, the anti-VEGF agent and phototherapy agentare administered to the skin, a tumor, surgical site, or a wound site.In an embodiment, for example, the anti-VEGF agent and phototherapyagent are administered and/or delivered to a blood vessel, lung, heart,throat, ear, rectum, bladder, stomach, intestine, esophagus, liver,brain, prostrate, breast or pancreas of the subject.

Dosages of the present opioids and structurally related derivatives willbe variable depending on many factors, including, for example, the routeof administration. If injected into the eye, for example, the dose willbe very small whereas oral administration will involve a larger dosagedue to transport of charged agent. A useful range for administered doseof the present therapeutic agents is 0.001 mg to 0.5 g. The formulationsfor treatment of macular degeneration comprise a phototherapy agent(Type I or II), and one or more VEGF inhibitors of this invention withor without additional starting base in normal saline, buffer, GRASincipients, etc. In some embodiments for the treatment of cancers byphototherapy, the phototherapy agent with the VEGF inhibitor and acytotoxic is administered. (−)R-naltrexone methobromide is synergisticwith 5-fluorouracil in endothelial cells and is also synergistic withAvastin to inhibit VEGF expression (the agents work differently).Recited cytotoxics for cancer therapy include a broad range of knownagents as combination therapy including, but not limited to,5-Fluorouracil, floxuridine, furtulon, capecitabine, gemcitabine, taxol(all forms), doxorubicin, cisplatin, and/or in conjunction with otherVEGF inhibitors like thalidomide or Avastin. The invention furtherincludes combination therapies wherein the present opioids andstructurally related derivatives are administered with one or more othertherapeutic agents and/or diagnostic agents, including, but not limitedto, alkylating agents, anti-metabolites, anti-cytoskeletal agents,topoisomerase inhibitors, anti-hormonal agents, targeted therapeuticagents, and the like. Examples of alkylating agents include, but are notlimited to, altretamine, benzodopa, busulfan, carboplatin, carboquone,carmustine, chlorambucil, chlornaphazine, chlorophosphamide,chlorozotocin, cisplatin, cyclosphosphamide, dacarbazine (DTIC),estramustine, fotemustine, ifosfamide, improsulfan, lomustine,mechlorethamine, mechlorethamine oxide hydrochloride, melphalan,meturedopa, nimustine, novembichin, phenesterine, piposulfan,prednimustine, ranimustine, temozolomide, thiotepa, triethylenemelamine,triethylenephosphoramide, triethylenethiophosphaoramide,trimethylolomelamine, trofosfamide, uracil mustard and uredopa. Suitableanti-metabolites include, but are not limited to aminopterin,ancitabine, azacitidine, 6-azauridine, capecitabine, carmofur,cytarabine or cytosine arabinoside (Ara-C), dideoxyuridine, denopterin,doxifluridine, enocitabine, floxuridine, fludarabine, 5-fluorouracil(5-FU), gemcetabine, leucovorin (folinic acid), 6-mercaptopurine,methotrexate, pemetrexed, pteropterin, thiamiprine, trimetrexate, andthioguanine. Examples of suitable anti-cytoskeletal agents include, butare not limited to, colchicines, docetaxel, macromycin, paclitaxel(taxol), vinblastine, vincristine, vindesine, and vinorelbine. Suitabletopoisomerase inhibitors include, but are not limited to, amsacrine,etoposide (VP-16), irinotecan, RFS 2000, teniposide, and topotecan.Examples of suitable anti-hormonal agents include, but are not limitedto, aminoglutethimide, aromatase inhibiting 4(5)-imidazoles,bicalutamide, finasteride, flutamide, goserelin, 4-hydroxytamoxifen,keoxifene, leuprolide, LY117018, mitotane, nilutamide, onapristone,raloxifene, tamoxifen, toremifene, and trilostane. Examples of targetedtherapeutic agents include, but are not limited to, a monoclonalantibodies such as alemtuzumab, bevacizumab, capecitabine, cetuximab,gemtuzumab, heregulin, rituximab, trastuzumab; tyrosine kinaseinhibitors such as imatinib and mesylate; and growth inhibitorypolypeptides such as erythropoietin, interleukins (e.g., IL-1, IL-2,IL-3, IL-6), leukemia inhibitory factor, interferons, thrombopoietin,TNF-α, CD30 ligand, 4-1BB ligand, and Apo-1 ligand. The (+) alkaloidbases also have potential activity as anti-inflammatory cytokineinhibitors. The combination of base with charged complement will actsynergistically for an indication like macular degeneration sincebenefit has been shown with other combo agents that include an anti-VEGFtherapeutic.

A range of phototherapy agents are useful in the present methods,including Type 1 and Type 2 phototherapy agents. Type 2 phototherapyagents for the present methods, include, but are not limited to,Photofrin®, Visudyne®, benzoporphyrins, phthalocyanines, phenothiazines,chlorins, bacteriochlorins, phthalocyanines, porphyrins, purpurins,merocyanines, pheophorbides, psoralens, aminolevulinic acid (ALA),hematoporphyrin derivatives, porphycenes, porphacyanine, and expandedporphyrin-like compounds and pro-drugs such as δ-aminolevulinic acid,which can produce drugs such as protoporphyrin. Type 1 phototherapyagents include, but are not limited to, cyanines, indocyanines,phthalocyanines, rhodamines, phenoxazines, phenothiazines,phenoselenazines, fluoresceins, squaraines, corrins, croconiums, azodyes, methine dyes, indolenium dyes, halogens, anthracylines, azides,C₁-C₂₀ peroxyalkyls, C₅-C₂₀ peroxyaryls, C₁-C₂₀ sulfenatoalkyls,sulfenatoaryls, azo dyes, naphthalocyanines, methylene blues, andchalcogenopyrylium analogues. Specific photosensitizers useful in someembodiments of the present invention are further described in any ofU.S. Pat. Nos. 7,303,926, 7,230,088, 7,198,778, 7,235,685; 7,011,817;7,201,892; 7,128,896; 6,485,704; 5,438,071; 5,405,957; 5,198,460;5,190,966; 5,173,504; 5,171,741; 5,166,197; 5,095,030; 5,093,349;5,079,262; 5,028,621; 5,002,962; 4,968,715; 4,920,143; 4,883,790;4,866,168; and 4,649,151. Photosensitizers useful in the presentinvention are further described in any of International PatentPublication Nos. WO 2003/032900, WO 2005/089813, and WO 2007/103250, andU.S. Patent Application Publication No. US 2002/0169107.

In some methods for phototherapy, the target region is illuminated withelectromagnetic radiation having wavelengths in the range of about 350nm to about 1300 nm, preferably for some applications in the range ofabout 400 nm to about 900 nm. In an embodiment, the target region isilluminated with electromagnetic radiation having wavelengths in therange of about 300 nm to about 900 nm. In some embodiments, thewavelength of the electromagnetic radiation corresponds to a peak in theabsorption spectrum of the phototherapy agent, for example is within 20nanometers of a peak in the absorption spectrum of the phototherapyagent in the visible or NIR regions. In some phototherapy procedures thetarget site is exposed to electromagnetic radiation having fluence,dosage and/or power sufficient to activate the phototherapy agent so asto induce cell death, for example via necrosis or apoptosis processes.In some embodiments, electromagnetic radiation having low energy (e.g.,less than 200 mW/cm² or optionally less than 100 mW/cm²), power orfluence, but sufficient dosage, is provided to activate the phototherapyagent without undesirable thermal effects. If the region of interest is,for example, a lesion or tumor on the skin surface, the region can bedirectly illuminated. In some methods, endoscopic and/or endoluminalcatheters can be employed to deliver electromagnetic radiation to thesubject to provide a photodiagnostic and/or phototherapeutic effect. Theintensity, power, and duration of the illumination and the wavelength ofthe electromagnetic radiation can vary widely depending on the bodylocation, the lesion site, the effect to be achieved, etc. In anembodiment, the power of the applied electromagnetic radiation ispreferably selected over the range of 1-500 mW/cm² and optionally forsome applications is selected over the range of 1-200 mW/cm² andoptionally for some applications selected over the range of 1-100mW/cm². In an embodiment, the duration of the exposure to appliedelectromagnetic radiation is selected over the range of 1 second to 60minutes, and optionally for some applications is selected over the rangeof 1 second to 30 minutes, and optionally for some applications isselected over the range of 1 second to 10 minutes, and optionally forsome applications is selected over the range of 1 second to 1 minute.

In embodiments, subjects of the invention can be any mammal, such as ahuman, and optionally the subject of the present methods is a patient inneed of treatment and/or diagnosis. The present methods are also usefulfor ex vivo and in vitro procedures, including medical therapeutic anddiagnostic procedures. As will be understood by one having skill in theart, the optical conditions for exposing the optical agent administeredto the patient to electromagnetic radiation will vary considerably withthe (i) therapeutic and/or diagnostic objectives, and (ii) the conditionof the subject (e.g., height, weight, state of health etc.). In anembodiment, the applied electromagnetic radiation has wavelengths,energy and/or fluence sufficient to achieve a desired therapeutic and/ordiagnostic result. In an embodiment, the electromagnetic radiation haswavelengths, energy and/or fluence sufficient to activate the opticalagent. Optionally, excitation is achieved using electromagneticradiation substantially free (e.g., less than about 10% of total radiantenergy), of ultraviolet radiation, for example, to minimize exposure ofthe subject to electromagnetic radiation capable of causing unwantedcell or tissue damage. Electromagnetic radiation can be provided to theoptical agent using a range of optical sources and/or surgicalinstrumentation, including a laser, electromagnetic radiation emittingdiodes, fiber optic device, endoscope, catheter, optical filters,mirrors, lenses, or any combination of these.

In an embodiment, the method further comprises contacting a targettissue of a patient in need of treatment with an effective amount of acompound of any one of formulae (FX1)-(FX164) and an effective amount ofa phototherapy agent. In a specific method, the target tissue is atumor, a region of abnormal tissue growth or region of abnormal bloodvessel growth. In a specific method, the target tissue is a mammarycarcinoma, an esophageal carcinoma, an endobronchial carcinoma, abladder tumor, a cervical tumor, a brain tumor, a head and neck tumor, alung tumor, a pituitary tumor, an intrathoracic tumor or a skinmalignancy. The present methods have broad clinical utility whichincludes, but is not limited to, phototherapy of tumors, inflammatoryprocesses, and impaired vasculature. In embodiments, subjects of theinvention may be any mammal, such as a human, and optionally the subjectof the present methods is a patient in need of treatment and/ordiagnosis. The present methods include in vivo, ex vivo and in vitroprocedures.

In an embodiment, a therapeutically effective amount of the anti-VEGFagent is provided to the subject. For example, parenteral administrationadvantageously contains a sterile aqueous solution or suspension of theanti-VEGF agent having a concentration of an active agent comprising thecompound of any one of formulae (FX1)-(FX164) ranging from about 0.01 μMto about 0.5M. Preferred parenteral formulations have a concentration ofan active agent comprising the compound of any one of formulae(FX1)-(FX164) selected over the range of 1 μM to 10 mM. Such solutionsalso may contain pharmaceutically acceptable buffers, emulsifiers,surfactants, and, optionally, electrolytes such as sodium chloride. Thedose of the compound of any one of formulae (FX1)-(FX164) may vary from0.1 to 500 mg/kg body weight, preferably from 0.5 to 2 mg/kg bodyweight.

In methods of the present invention, the anti-VEGF agent and/orphototherapy agent can be formulated for enteral (oral or rectal),parenteral, topical, or cutaneous administration. Topical or cutaneousdelivery of the anti-VEGF agent and/or phototherapy agent may alsoinclude aerosols, creams, gels, solutions, emulsions and colloids. Thecompositions are administered in doses effective to achieve the desireddiagnostic or therapeutic objective. Such doses may vary widelydepending upon the particular complex employed, the organs or tissues tobe examined or treated, the equipment employed in the clinicalprocedure, the efficacy of the treatment achieved, and the like. Thesecompositions contain an effective amount of the anti-VEGF agent andphototherapy agent along with conventional pharmaceutical carriers andexcipients appropriate for the type of administration contemplated.These compositions may also include stabilizing agents and skinpenetration enhancing agents and also may contain pharmaceuticallyacceptable buffers, emulsifiers, surfactants, and, optionally,electrolytes such as sodium chloride. Formulations for enteraladministration may vary widely, as is well known in the art. In general,such formulations are liquids, which include an effective amount of thecomplexes in aqueous solution or suspension. Such enteral compositionsmay optionally include buffers, surfactants, emulsifiers, thixotropicagents, and the like. Compounds for oral administration may also containflavoring agents and other ingredients for enhancing their organolepticqualities. Formulations for topical delivery may also contain liquid orsemisolid excipients to assist in the penetration of thephotosensitizer. The compounds may also be delivered in an aerosolspray.

As will be appreciated by one having skill in the art, the fluenceemployed during excitation of the phototherapy agent can vary dependingon the type of tissue, depth of the target, composition of thephototherapy agent and the amount on composition of overlying fluid andblood. In some embodiments, the fluence employed is selected over therange of 10 to 500 Joules/cm⁻². The irradiance is typically selectedfrom the range of 50 to 1500 mW/cm⁻², preferably of 50 to 500 mW/cm⁻².Electromagnetic radiation may be provided to the phototherapy agentusing a range of optical sources and/or surgical instrumentation,including a laser, light emitting diodes, fiber optic device, endoscope,catheter, optical filters, or any combination of these.

Example 2 Methods of Using Anti-VEGF Opioids and Structurally RelatedDerivatives for Treatment and Management of Macular Degeneration

Macular degeneration (MD) is an eye disease that destroys central visionby damaging the macula, a thin layer of nerve cells that lines most ofthe inside of the eye. There are two forms of MD: atrophic (“dry”) andexudative (“wet”). About ninety percent of patients have the dry form;whereas, only ten percent have the “wet” form. Patients with the wetform can lose up to ninety percent of their vision rapidly. Maculardegeneration is the most common cause of visual loss in individuals overthe age of 60 and has destroyed the central vision of 1.7 millionAmericans with another 11 million at risk. 200,000 new cases ofadvanced, age-related macular degeneration (AMD) are identified eachyear in the United States. Currently, there is no known cure. Since theelderly is the fastest growing segment of the population, MD will becomea major economic and social problem. Thus, there is an urgent need foreffective treatments for MD, particularly AMD

Wet macular degeneration develops when new blood vessels grow from thechoroid underneath the macular portion of the retina. These new vesselsare called choroidal neovascularizations (CNVs). These vessels leakfluid or blood, thus the term “wet” macular degeneration. Much like thedry form of macular degeneration, a breakdown in the vasculararchitecture activates the abnormal growth of blood vessels. Withoutnormal perfusion of the macula, the tissue deteriorates. Laserphotocoagulation was the only routine therapeutic option used for wet MDuntil recently and provided only modest response. Less intensephototherapy using the photoreactive dye, Verteporfin, has recently beenused to treat wet MD. Verteporfin is a blood-vessel-blocking agent thatis administered via injection. Activated verteporfin generates highlyreactive oxygen species resulting in local damage to neovascularendothelium. This causes vessel occlusion and reduced progression of thedisease. However, since verteporfin can accumulate in the retina,collateral damage to retinal structures may unfortunately result withconcomitant loss of some remaining vision.

The first antiangiogenic agent, α-interferon, clinically tested for MDwas ineffective and resulted in a high rate of adverse effects. A numberof other antiangiogenic drugs are in development, including angiostaticsteroids (e.g., anecortave acetate, Alcon) and antibodies or antibodyfragments of vascular epidermal growth factor (VEGF) (e.g.Avastin-bevacizumab). Additional new drugs for the treatment of MDinclude EYE101 (Eyetech Pharmaceuticals), LY333531 (Eli Lilly), andRETISERT implant (Bausch & Lomb), which exudes a steroid into the eyefor up to three years. The most successful currently FDA approved agentis Lucentis (ranibizumab), a humanized anti-VEGF antibody fragment thatinhibits VEGF activity by competitive binding. Lucentis is derived fromAvastin (bevacizumab), a full-length humanized monoclonal antibodyagainst VEGF. About 95% patients with wet macular degeneration maintaintheir baseline vision after 12 months of treatment with Lucentis. Bothagents are given by injection directly into the affected eye(s) everyonce a month for the first four months then every three months for therest of the patient's life. The downside of Lucentis® and Avastin®treatment is that they stop the abnormal vessels from growing andleaking but often don't cause permanent closure. Therefore, injectionsof either drug have to be given repeatedly. PDT can close vesselspermanently but often at the price of vision loss. Combination therapyis most efficacious. The most common combination now used is PDT withhalf the amount of laser dose followed with an injection of Lucentis® orof Avastin® and optionally with an anti-inflammatory steroid.

The present invention provides opioids and structurally relatedderivatives useful as anti-angiogenesis agents and anti-VEGF agents forin treatment of ocular neovascular diseases, including maculardegeneration such as age related macular degeneration. In someembodiments, for example, the present invention provides methodscomprising in vivo administration of an anti-VEGF agent comprising anopioid or structurally related derivative to a patient in need oftreatment, optionally further including administration and excitation ofa phototherapy agent. FIG. 3 provides flow diagrams illustrating methodsof the present invention for the treatment of ocular neovasculardiseases including the step of administering an anti-VEGF agent. Asshown in this figure, the present methods and compositions for treatingocular neovascular disease encompass both monotherapies and combinationtherapies in conjunction with administration and excitation of aphototherapy agent. In the direct monotherapy, an anti-VEGF agent isadministered to a target tissue. Administration results in suppression,alleviation and/or reduction of unwanted angiogenesis, thereby resultingin treatment or prevention of an ocular neovascular disease, such as MDand AMD. In the combination therapy, an anti-VEGF agent and phototherapyagent are co-administered to a target tissue. Administration followed byactivation results in treatment or prevention of ocular neovasculardisease wherein the anti-VEGF agent suppresses, alleviates and/orreduces unwanted angiogenesis.

Anti-VEGF and Anti-angiogenic agents of the methods of this aspect ofthe invention include a large number of opioid antagonists, opioidagonists and structurally related derivatives thereof including, but notlimited to, quaternary salts and isomeric and epimeric forms of(+/−)R/S-Naloxone, (+/−)R/S-Nalmefene, nalbuphine, (+/−)-oxymorphone,(+/−)-hydrocodone, (+/−)-oxycodone, (+/−)-hydromorphone,tetrahydrosinomenine, dextrorphan, and dextromethorphan. These compoundsprovide anti-VEGF therapeutic agents effective for treatment of ocularneovascular disease.

In an embodiment, the invention provides a method of treating an ocularneovascular disease, the method comprising administering to a patient aneffective amount of a compound having the formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═O)—, —(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶)—; each of R¹ and R²is independently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl,C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; each of R³-R¹⁶ and R⁵⁰-R⁵⁴ is independently—H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₂-C₈alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀ hydroxyalkyl, orC₅-C₁₀ carbonylalkenylheteroaryl; and n is 0 or 1, wherein: when n is 1,the nitrogen to which R² is attached has a positive charge andoptionally is associated with an anion; when n is 0, R² is not present;and when in (FX1) each of X and Y is —OH, n is 1, and W is —(C═O)—, theneach of R¹ and R² is independently a group other than cyclopropylmethyl,and each of R¹ and R² is independently a group other than methyl. In anembodiment, the invention provides a method of treating an ocularneovascular disease, the method comprising administering to a patient aneffective amount of a compound having the formula (FX61):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H,or —OH;

each of R¹ and R² is independently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀carbonylalkenylaryl, or C₅-C₁₀ carbonylalkenylheteroaryl; each ofR²⁰-R²⁹ is independently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl,C₂-C₈ alkenyl, C₁-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl,C₅-C₁₀ carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl; and n is 0 or1, wherein: when n is 1, the nitrogen to which R² is attached has apositive charge and optionally is associated with an anion; and when nis 0, R² is not present. In an embodiment, the method of this aspectfurther comprises (i) administering to a patient an effective amount ofa phototherapy agent and (ii) exposing the phototherapy agentadministered to the patient to electromagnetic radiation. In anembodiment, the method of this aspect of the invention comprisesadministering to a patient a compound having any one of formulae(FX1)-(FX164), including all of the specific composition classes andcompounds described herein, in combination with any of the method stepspresented herein. In an embodiment, the method of this aspect of theinvention comprises administering to a target tissue of a patient acompound having any one of formulae (FX1)-(FX164), including all of thespecific composition classes and compounds described herein, incombination with any of the method steps presented herein. As will beunderstood by one of skill in the art, the present methods expresslyinclude administering one or more anti-VEGF agents, including thecompound classes, compounds, and variations thereof, described herein,including the compound classes, compounds and variations described inconnection with any one of formulae (FX1)-(FX164), optionally incombination with co-administering and activating a phototherapy agent.

In an embodiment, the ocular neovascular disease is age related maculardegeneration. In an embodiment, the ocular neovascular disease isischemic retinopathy, intraocular neovascularization,neovascularization, retinal neovascularization, choroidalneovascularization, diabetic macular edema, diabetic retina ischemia,diabetic retinal edema or proliferative diabetic retinopathy. In anembodiment, a method of this aspect further includes contactingneovascular endothelium cells of a patient undergoing treatment with acompound having any one of formulae (FX1)-(FX164), and optionallycontacting neovascular endothelium cells of a patient undergoingtreatment with a phototherapy agent. In some embodiments, administrationof a compound having any one of formulae (FX1)-(FX164) or pharmaceuticalformulation thereof is delivered to an eye tissue of the patient; andoptionally wherein delivery is via injection or topical delivery.

Example 3 Methods of Using Anti-VEGF Opioids and Structurally RelatedDerivatives for Treatment and Management of Cancer

Opioids are a group of drugs that exhibit opium or morphine-likeproperties. They are employed primarily as moderate to stronganalgesics, but have many other pharmacological effects as well,including drowsiness, respiratory depression, constipation, changes inmood, and mental clouding without a resulting loss of consciousness.Morphine and codeine are by far the most important naturally occurringopiate agonists. Substitution at the nitrogen atom of the morphinan ringof these compounds affects their pharmacologic properties. For example,morphinan derivatives having various nitrogen substituents exhibitpartial agonist/antagonist activity or potent antagonist activity.Morphinan compounds comprising quaternary amines are also known. Forexample, some N-oxides of certain morphinans have been variouslyreported to be less active than the corresponding tertiary amine.

Recent research has shown that morphinans comprising opiate receptorantagonists and, in particular, their respective quaternary amines arepotent inhibitors of vascular endothelial growth factor (VEGF). VEGFinhibitors are important adjuncts in the treatment of various tumors andthe treatment of macular degeneration. More recent work has suggestedthat VEGF inhibitory activity appears to be independent of thestereochemistry of the morphinan ring system. That is, (+)-morphinanquaternary compounds appear to inhibit VEGF as well as thepharmacologically active (−)-morphinan quaternary compounds. Thus, other(+)-morphinan derivatives such as (+)-morphinanium N-oxides may beuseful as improved VEGF inhibitors because they do not interact withopiate receptors.

The invention provides methods of treating cancer by administration of atherapeutically effective amount of an opioid compound, structurallyrelated opioid derivative or isomer thereof. In an embodiment, forexample, the invention provides a method of treating cancer, the methodcomprising administering to a patient in need of treatment an effectiveamount of a compound having the formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₅ alkoxy;

Y is —H or —OH; W is —(CR¹⁰R¹¹)—, —(C═S)—, or —(CR¹⁴NR¹⁵R¹⁶)—; each ofR¹ and R² is independently —H, —CH₃, —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀carbonylalkenylaryl, or C₅-C₁₀ carbonylalkenylheteroaryl; each of R¹⁰and R¹¹ is independently —OCH₃, C₁-C₈ alkyl, C₃-C₃ cycloalkyl, C₂-C₈alkenyl, C₂-C₈ alkoxy, C₂-C₈ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl,C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀hydroxyalkyl, or C₅-C₁₀ carbonylalkenylheteroaryl; each of R³-R⁹,R¹²-R¹⁶, and R⁵⁰-R⁵⁴ is independently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈cycloalkyl, C₂-C₅ alkenyl, C₂-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl,C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈haloalkyl, C₁-C₁₀ hydroxyalkyl, or C₅-C₁₀ carbonylalkenylheteroaryl; andn is 0 or 1, wherein: when n is 1, the nitrogen to which R² is attachedhas a positive charge and optionally is associated with an anion; andwhen n is 0, R² is not present. In an embodiment, for example, theinvention provides a method of treating cancer, the method comprisingadministering to a patient in need of treatment an effective amount of acompound having the formula (FX1) or (FX2):

wherein: ring A is

X is —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H or —OH; W is —(CR¹⁰R¹¹)—,—(C═O)—, —(C═S)—, —(C═CR¹²R¹³)—, or —(CR¹⁴NR¹⁵R¹⁶); R¹ is —H, —CH₃,—(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; and each of R³-R¹⁶ and R⁵⁰-R⁵⁴ isindependently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈alkenyl, C₂-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl,C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₁-C₁₀hydroxyalkyl, or C₅-C₁₀ carbonylalkenylheteroaryl. In an embodiment, forexample, the invention provides a method of treating cancer, the methodcomprising administering to a patient in need of treatment an effectiveamount of a compound having the formula (FX61):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H,or —OH;

each of R¹ and R² is independently —(O⁻), C₁-C₁₀ alkyl, C₃-C₁₀cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀ haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀ alkylheteroaryl, C₅-C₁₀carbonylalkenylaryl, or C₅-C₁₀ carbonylalkenylheteroaryl; each ofR²⁰-R²⁹ is independently —H, —OH, —OCH₃, C₁-C₈ alkyl, C₃-C₅ cycloalkyl,C₂-C₈ alkenyl, C₁-C₈ alkoxy, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀alkylaryl, C₅-C₁₀ alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl,C₅-C₁₀ carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl; and n is 0 or1, wherein: when n is 1, the nitrogen to which R² is attached has apositive charge and optionally is associated with an anion; and when nis 0, R² is not present. In an embodiment, for example, the inventionprovides a method of treating cancer, the method comprisingadministering to a patient in need of treatment an effective amount of acompound having the formula (FX61):

wherein: each X is independently —OH, —OCH₃, or C₂-C₈ alkoxy; Y is —H,or —OH;

R¹ is —H, —CH₃, C₁-C₁₀ alkyl, C₃-C₁₀ cycloalkyl, C₂-C₁₀ alkenyl, C₁-C₁₀haloalkyl, C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, C₅-C₁₀ carbonylalkenylaryl, or C₅-C₁₀carbonylalkenylheteroaryl; and each of R²⁰-R²⁹ is independently —H, —OH,—OCH₃, C₁-C₈ alkyl, C₃-C₈ cycloalkyl, C₂-C₈ alkenyl, C₁-C₈ alkoxy,C₅-C₁₀ aryl, C₅-C₁₀ heteroaryl, C₅-C₁₀ alkylaryl, C₅-C₁₀alkylheteroaryl, halo, nitrile, C₁-C₈ haloalkyl, C₅-C₁₀carbonylalkenylheteroaryl, or C₁-C₁₀ hydroxyalkyl. In an embodiment, themethod of treating cancer of this aspect of the invention comprisesadministering to a patient a compound having any one of formulae(FX1)-(FX164), including all of the specific compositions classes andcompounds described herein, in combination with any of the method stepspresented herein. In an embodiment, the method of treating cancer ofthis aspect of the invention comprises administering to a target tissueof a patient a compound having any one of formulae (FX1)-(FX164),including all of the specific composition classes and compoundsdescribed herein, in combination with any of the method steps presentedherein. As will be understood by one of skill in the art, the presentmethods of treating cancer expressly include administering one or moreanti-VEGF agents including the compound classes, compounds, andvariations thereof, described herein, including the compound classes,compounds and variations described in connection with any one offormulae (FX1)-(FX164), optionally in combination with co-administeringand activating a phototherapy agent.

In an aspect, the invention provides methods of attenuating migrationand/or proliferation of endothelial cells of a tumor or cancer,comprising contacting the cells with a therapeutically effective amountof an opioid compound, structurally related opioid derivative or isomerthereof, such as a compound having any of formulae (FX1)-(FX164). Inanother aspect, the invention provides a method of treating abnormalangiogenesis associated with cancer, comprising administering to atarget tissue, such as cancer tissue or a tumor, of the patient atherapeutically effective amount of an opioid compound, structurallyrelated opioid derivative or isomer thereof, such as a compound havingany of formulae (FX1)-(FX164), in an amount which is effective to atleast partially attenuate abnormal angiogenesis. The invention alsoprovides a method of treating abnormal neovascularization associatedwith cancer, comprising administering to a target tissue, such as cancertissue or a tumor, of a patient in need of such treatment, atherapeutically effective amount of an opioid compound, structurallyrelated opioid derivative or isomer thereof, such as a compound havingany of formulae (FX1)-(FX164), effective to at least partially inhibitlocal formation of blood vessels. The invention provides a method ofattenuating proliferation of hyperproliferative cells associated withcancer in a subject, comprising administering to a target tissue, suchas cancer tissue or a tumor, of the subject at least one opioidcompound, structurally related opioid derivative or isomer thereof, suchas a compound having any of formulae (FX1)-(FX164), in an amount whichis effective to attenuate proliferation of the hyperproliferative cells.

In an aspect, the invention provides methods to treat or inhibit theonset, development or recurrence of cancer. The invention provides amethod of attenuating tumor progression and metastasis in animaltissues, comprising contacting a target tissue, such as tumor cells ortissues, with a therapeutically effective amount of an opioid compound,structurally related opioid derivative or isomer thereof, such as acompound having any of formulae (FX1)-(FX164). In another aspect, theinvention provides a method of reducing the risk of recurrence of acancer or tumor after medical intervention (such intervention toinclude, but not be limited to surgery, e.g. pulmonary surgery, surgicaland endoscopic procedures, e.g. colonoscopy, gastrolaparoscopy,chemotherapy, etc.), comprising administering to a cancer patient anopioid compound, structurally related opioid derivative or isomerthereof, such as a compound having any of formulae (FX1)-(FX164).Therefore, as will be apparent to one having skill in the art, theinvention contemplates, for example, a method of minimizingpost-operative recurrence of cancer in a patient, comprisingadministering to a cancer patient an opioid compound, structurallyrelated opioid derivative or isomer thereof, such as a compound havingany of formulae (FX1)-(FX164). In an embodiment of the invention, thecancer is prostate cancer, gastrointestinal cancer, colon cancer,colorectal cancer, pancreatic cancer, breast cancer, lung cancer, throatcancer, skin cancer, stomach cancer, pancreatic cancer, brain cancer,liver cancer, prostate cancer, kidney cancer, bladder cancer, bonecancer, brain cancer, eye cancer, gallbladder cancer, head and neckcancer, Hodgkin lymphoma, mouth cancer, ovarian cancer, testicularcancer, throat cancer, esophageal cancer, small intestine cancer,pharyngeal cancer, laryngeal cancer, urethral cancer, uterine cancer,parathyroid cancer, cervical cancer, penile cancer or vaginal cancer.The compounds of the present invention may also be useful for thetreatment of cancer in patients, as described above, either when usedalone or in combination with one or more other anticancer agents, e.g.,radiotherapy and/or other chemotherapeutic, including antiangiogenic,treatments conventionally administered to patients for treating cancer.Several main categories and examples of such drugs are listed herein andinclude, but are not limited to metalloproatease inhibitors, inhibitorsof endothelial cell proliferation/migration, antagonists of angiogenicgrowth factors, inhibitors of Integrin/Survival signaling, and chelatorsof copper.

The compositions and methods of the present invention may be oftherapeutic value in treatment for patients who have cancers and/ortumors that develop and/or progress (e.g., grow, become invasive,metastasize, etc.), at least in part, via one or more angiogenesisprocesses. Cancers and tumors that develop and/or progress, at least inpart, via one or more angiogenesis processes include carcinomas,sarcomas, leukemias, blastoma, myelomas, and solid tumors. Cancers andsolid tumors include, but are not limited to, adrenal corticalcarcinoma, tumors of the bladder: squamous cell carcinoma, urothelialcarcinomas; tumors of the bone: adamantinoma, aneurysmal bone cysts,chondroblastoma, chondroma, chondromyxoid fibroma, chondrosarcoma,fibrous dysplasia of the bone, giant cell tumour, osteochondroma,osteosarcoma; breast tumors: secretory ductal carcinoma, chordoma; colontumors: colorectal adenocarcinoma; eye tumors: posterior uveal melanoma,fibrogenesis imperfecta ossium, head and neck squamous cell carcinoma;kidney tumors: chromophobe renal cell carcinoma, clear cell renal cellcarcinoma, nephroblastoma (Wilms tumor), kidney: papillary renal cellcarcinoma, primary renal ASPSCR1-TFE3 tumor, renal cell carcinoma; livertumors: hepatoblastoma, hepatocellular carcinoma; lung tumors: non-smallcell carcinoma, small cell cancer; malignant melanoma of soft parts;nervous system tumors: medulloblastoma, meningioma, neuroblastoma,astrocytic tumors, ependymomas, peripheral nerve sheath tumors,phaeochromocytoma; ovarian tumors: epithelial tumors, germ cell tumors,sex cord-stromal tumors, pericytoma; pituitary adenomas; rhabdoid tumor;skin tumors: cutaneous benign fibrous histiocytomas; smooth muscletumors: intravenous leiomyomatosis; soft tissue tumors: liposarcoma,myxoid liposarcoma, low grade fibromyxoid sarcoma, leiomyosarcoma,alveolar soft part sarcoma, angiomatoid fibrous histiocytoma (AFH),clear cell sarcoma, desmoplastic small round cell tumor, elastofibroma,Ewing's tumors, extraskeletal myxoid chondrosarcoma, inflammatorymyofibroblastic tumor, lipoblastoma, lipoma/benign lipomatous tumors,liposarcoma/malignant lipomatous tumors, malignant myoepithelioma,rhabdomyosarcoma, synovial sarcoma, squamous cell cancer; tumors of thetestis: germ cell tumors, spermatocytic seminoma; thyroid tumors:anaplastic (undifferentiated) carcinoma, oncocytic tumors, papillarycarcinoma; uterus tumors: carcinoma of the cervix, endometrialcarcinoma, leiomyoma and the like.

The compositions and methods of the present invention may also be oftherapeutic value in treatment for cancer patients who havenonangiogenic cancers and tumors, as many of these types of cancertumors are known to undergo spontaneous transformation into anangiogeneic phenotype under some conditions. Cancers not involvingangiogenesis include those that do not involve the formation of a solidtumor fed by neovasculature. Certain blood cell cancers can fall intothis category, for example: leukemias, including acute lymphocyticleukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocyticleukemia (CLL), chronic myelogenous leukemia (CML), and hairy cellleukemia; lymphomas (arising in the lymph nodes or lymphocytes)including Hodgkin lymphoma, Burkitt's lymphoma, cutaneous lymphoma,cutaneous T-cell lymphoma, follicular lymphoma, lymphoblastic lymphoma,MALT lymphoma, mantle cell lymphoma, Waldenstrom's macroglobulinemia,primary central nervous system lymphoma; and some cancers of the bonemarrow elements including Ewing's sarcoma and osteosarcoma. Thus, in oneaspect, the invention provides a method of treating cancer, wherein thecancer may be associated with an angiogenesis process or may be notassociated with angiogenesis process.

Therapeutic methods for treatment of ocular neovascular disease and someforms of cancer can optionally include additional method steps,processes, variations, compositions, phototherapy agents, formulations,delivery conditions and phototherapeutic conditions as described abovein connection with the phototherapy methods of the present invention.

Example 4 Quaternary Alkaloid VEGF Inhibitors as Adjuncts for theTreatment of Cancer and Macular Degeneration and in Phototherapy

The invention provides compounds, such as morphinanium compounds andrelated derivative, salts and alkaloid forms, that inhibit metastasisinduction by activation of the inflammatory response to excessivereactive oxygen species produced during photodynamic therapy (PDT) andother angiogenesis initiating therapies and disease conditions.

For example, it is believed that certain Type II Photodynamic therapy(PDT) agents initiate the up-regulation of VEGF that then promotetumorigenesis in certain vascular cancer types. Accelerated diseasepresents as gross metastasis. VEGF inhibitors effectively reduce thelevels of VEGF post-PDT and thus mitigate side-effects andproliferation. (−)-R-Naltrexone methobromide (MNTX) may provide a potentVEGF inhibitor in some therapies. For example, MNTX may attenuatevascular permeability due to disease, injury or cancer. For example,MNTX may act synergistically with 5-fluorouracil and bevacizumab insuppressing VEGF-induced human pulmonary microvascular endothelial cell(EC) proliferation and migration, which are two key components incancer-associated angiogenesis. For example, MNTX provides an activeinhibitor of pancreatic cancer in mouse models. The synergistic effectof VEGF antibody, PDT/Typell, Gemcitabine delivered by a non-targetednanoparticle/liposomal methodology, significantly reduces pancreatictumor weight relative to controls. Because the structurally unrelatedquaternary alkaloid, berberine, also exhibits some inhibition of VEGFexpression, opiate receptor binding may not be a necessary criteria foradjunct or therapeutic candidacy. Sufficiency for activity is also metby the (+) enantiomer and other isomers of MNTX as well as other relatedquaternary alkaloid salts and the nitrogen oxides of (+ or −) ofmorphinan alkaloids.

To evaluate the effectiveness of the present compounds in the context oftherapeutic applications for the treatment of cancer and maculardegeneration and for use in a phototherapy procedure, in vitro assaytechniques such as cell proliferation and migration assay and VEGF ELISAmethods may be used.

4.1 In Vitro Assay Techniques for Determining Anti-VEGF Activity

Angiogenesis or the formation of new blood vessels is used by neoplasmto harness nutrients for their survival and growth. Endothelia cells areactivated by the nearby neoplastic cells that lead to secretion of VEGF,MMPS and other angiogenic stimuli. MMPS degrades the extracellularmatrix and VEGF stimulates angiogenesis. [See, (1) Folkman, J.Angiogenesis in cancer, vascular, rheumatoid and other diseases. Nat.Med., 1: 27-33, 1995; and (2) J. Folkman, The role of angiogenesis intumor growth, Cancer Biol., 1992, 3, 65-71]. As a result the endothelialcells invade the ECM where they migrate, proliferate and form new bloodvessels, which supports neoplasm growth and survival. The angiogenicpotential of neoplasm also correlates with its metastases potential.Hence by inhibiting some of these factors the angiogenesis process canbe stopped suppressing the tumor growth and metastases. [See, e.g.,Dvorak, H. F., Brown, L. F., Detmar, M., and Dvorak, A. M. Vascularpermeability factor/vascular endothelial growth factor, microvascularhyperpermeability and angiogenesis. Am. J. Pathol., 146: 1029-1039,1995.]

Cell Proliferation and Migration Assay

Proliferation and migration of Endothelial Cells (EC) is an importanthallmark of angiogenesis. [See. e.g., Folkman, J. Angiogenesis incancer, vascular, rheumatoid and other diseases. Nat. Med., 1: 27-33,1995]. Hence it is important to screen compounds for their inhibitorproperties on cell migration and proliferation. Human umbilical veinendothelial cells (HUVECs) and Human Pulmonary Microvascular EC(HPMEC)may be used to assess the anti-proliferative and migrating properties ofthe present compounds.

The cell migration assay is based on the cell migration in a modifiedBoyden chamber. [See, e.g., Lingen, M. W., 2002. Endothelial CellMigration Assay: A Quantitative Assay for Prediction of In Vivo Biology.Humana Press, Inc., pp. 337-347]. The upper chamber (UC) contains cellis treated with various test compounds and the lower chamber (LC)contains the agonist/chemo attractant. At the end of the assay,typically 24 hours, a cell proliferation reagent like WSt-1 is added.After 2 hours the absorbance of the cells is read with a plate reader.Any reduction in the absorbance values by the present compounds versusthat of control suggests that the compound has anti-angiogenicproperties.

The cell proliferation assay measures the ability of various compoundsof the invention to inhibit the growth of EC in presence VEGF orMorphine Sulfate. [See, e.g., Methylnaltrexone inhibits opiate andVEGF-induced angiogenesis: Role of receptor transactivation. P. A.Singleton et al./Microvascular Research 72 (2006) 3-11]. Briefly, cellsare pretreated with the test compounds followed by VEGF incubation.Negative controls are not pretreated and VEGF is added directly. Afterthe VEGF treatment, a cell proliferation reagent, such as WST-1, isadded and the absorbance is measured. Any reduction in absorbance valuesobserved for cells pre-treated with the present compounds versus nopre-treatment points toward the anti-angiogenic properties of thecompounds tested.

VEGF ELISA

VEGF is a potent, multifunctional, and endothelial cell specific growthfactor. The production of vascular endothelial growth factor (VEGF) bytumor cells plays an integral role in controlling proliferation andmetastasis. [See, e.g., Dvorak, H. F., Brown, L. F., Detmar, M., andDvorak, A. M. Vascular permeability factor/vascular endothelial growthfactor, microvascular hyperpermeability and angiogenesis. Am. J,Pathol., 146: 1029-1039, 1995]. It stimulates vasodilatation and cellproliferation, increases permeability and migration, and promotesendothelial cell survival. [See, e.g., Ferrara N: Vascular endothelialgrowth factor as a target for anticancer therapy. Oncologist 9: 2-10,2004]. It is upregulated in response to hypoxic conditions in the tumor[See, e.g., Pugh C W and Ratcliffe P J: Regulation of angiogenesis byhypoxia: Role of the HIF system. Nat Med 9: 677-684, 2003], chemotherapyand radiation induced stress [See, e.g., (1) Effect of chemotherapeuticstress on induction of vascular endothelial growth factor family membersand receptors in human colorectal cancer cells Mol Cancer Ther 2008;7(9):3064-70], (2) Targeting Chemotherapy-induced VEGF Up-regulation byVEGF Antisense Oligonucleotides in HNSCC Cell Lines. FRANK RIEDEL, KARLGÖTTE, ULRICH GOESSLER, HANNEN SADICK and KARL HÖRMANN. AnticancerResearch Jul. 1, 2004 vol. 24 no. 4 2179-2184, (3) Z-360, a NovelCholecystokinin-2/Gastrin Receptor Antagonist, InhibitsGemcitabine-Induced Expression of the Vascular Endothelial Growth FactorGene in Human Pancreatic Cancer Cells. Nobuyoshi KOBAYASHI, KoichiSETO,* Yuki ORIKAWA, Hiroki HAMANO, Koji YOSHINAGA, and Mineo TAKEI.Biol. Pharm. Bull. 33(2) 216-222 (2010)], and photodynamic therapy.[See, e.g., Effect of hypericin-mediated photodynamic therapy on theexpression of vascular endothelial growth factor in human nasopharyngealcarcinoma. INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 20: 421-428,2007].

The VEGF ELISA is an in vitro method that measures VEGF content inconditioned media, tissue lysates, cell lysates, and serum. Briefly,cells are pre-treated with compounds/drugs that induce VEGF expressionand its secretion in conditioned media. The conditioned media is thencollected and the secreted VEGF can be captured using a captureantibody. [See, e.g., Z-360, a Novel Cholecystokinin-2/Gastrin ReceptorAntagonist, Inhibits Gemcitabine-Induced Expression of the VascularEndothelial Growth Factor Gene in Human Pancreatic Cancer Cells.Nobuyoshi KOBAYASHI, Koichi SETO,* Yuki ORIKAWA, Hiroki HAMANO, KojiYOSHINAGA, and Mineo TAKEI. Biol. Pharm. Bull. 33(2) 216-222 (2010)].The captured VEGF is then detected as summarized in the general ELISAprotocol. (R&D Systems, Minneapolis, Minn.).

The present compounds are screened in a similar fashion for their effecton the VEGF levels in conditioned media. Cells are pretreated with thepresent compounds, for example for 1 hour, before actual treatment withVEGF inducing drugs/compounds. After a pre-determined treatment time,the conditioned media is collected and stored at −20 C. The cell lysateis also prepared for measuring the intracellular level of VEGF. The VEGFlevels are measured using the VEGF ELISA kit from R&D Systems(Minneapolis, Minn.). The VEGF levels are then be compared for anyreduction seen with pre-treatment of cells with the present compounds ascompared to cells without any pre-treatment.

In vitro cell proliferation and migration assay may be carried out inconnection with the compounds of the present invention. For example,such an assay approach may be carried out with respect to the fourrelated alkaloids shown below provided in a substantially purified formwith respect to other stereoisomers:

4.2 In Vivo Techniques for with Mouse Tumor Model

To demonstrate the effectiveness of the present compounds and methodsfor treatment of cancer and in phototherapy applications, in vivo mousetumor model experiments were carried in connection with the compoundMNTX, for example the purified stereoisommer (−)-R-naltrexonemethobromide. These experiments include comparative studies andpancreatic tumor growth inhibition studies.

Comparative Studies

In these experiments nude mice weighing approximately 25 grams wereused. First, the tail of the pancreas was exposed through an abdominalincision. Then 1×10⁶ AsPC-1 cells were injected into the pancreas. Thepancreas was replaced in the peritoneal cavity and the incision wasclosed. A 3-5 mm diameter tumor developed in 10 days.

MNTX was dissolved at 40 mg/mL in phosphate buffered saline (PBS) andthen formulated to 100 uL (50 uL drug and 50 uL PBS) and injected subQclose to the pancreas. One injection per day was given for 10 days. Thetotal dose was therefore 20 mg/mouse. For control and comparison,experiments on mice that were 1) not treated, 2) treated with lightalone, 3) treated with gemcitibine alone, and 4) treated with Avastinalone using essentially the same procedure as described above wereperformed. The mice were sacrificed at day 24 and tumor weight wasdetermined.

FIG. 4 provides local tumor burden experimental results for comparativestudies involving administration of MNTX, Avastin and Gemcitabineanti-tumor agents. Tumor weight (mg) is shown for tumor conditions of:no treatment (NT); treatment with light alone (light alone); treatmentwith Gemcitabine alone (GEM); treatment with Avastin alone (Avastin) andtreatment with MNTX (MNTX). FIG. 4 shows the tumor weight measured atsacrifice. As shown in FIG. 4, the treatment with MNTX was measurablysuperior to the control, Gemcitabine and Avastin conditions.

Pancreatic Tumor Growth Inhibition Studies

In these experiments, the effect of a purified stereoisomer of MNTX onAsPC-1 pancreatic cancer was evaluated. Specifically, the experimentsdemonstrate the potential application of (−)-R-naltrexone methobromide(abbreviated MNTX1) administration for treatment of pancreatic cancer.

The MNTX1 dose was established using an escalating dose study on mice.An intraperitoneal injection dose of 150 mg/kg was chosen. For a dosageof 400 mg/kg, 2 of 2 died, for a dosage of 200 mg/kg, 1 of 4 died; andfor a dosage of 150 mg/kg (300 μl of 10 mg/ml), 3 of 3 survived.Therefore, a dose of 150 mg/kg (Intraperitoneal injection, I.P.) wasused for each administration in the following orthotopic pancreatictumor growth inhibition experiments.

Nude mice weighing approximately 25 grams were used. The tail of thepancreas was exposed through an abdominal incision. Then 1×10⁶ AsPC-1cells were injected into the pancreas. The pancreas was replaced in theperitoneal cavity and the incision was closed. A 3-5 mm diameter tumordeveloped in 10 days.

MNTX1 was formulated at 10 mg/mL in PBS. The 150 mg/kg dose wasaccomplished by a 30 uL injection of a 10 mg/mL stock solution.Injections were done daily for 13 days, and the 10 animals weresubsequently sacrificed on day 24. Treatments start after 10 days ofAsPC1 pancreatic tumor cell orthotopic implantation. After two moreweeks, the mice are sacrificed and tumor weight is measured for theevaluation of tumor growth inhibition effects. For control, four micewere injected according to the above schedule with PBS only. They werealso sacrificed on day 24. FIG. 5 provides a schematic illustrating theexperimental design of the pancreatic tumor growth inhibition studies.I.P. treatments (150 mg MNTX1/kg/each injection, 300 μl of 10 mg/ml foreach injection totally for 13 times, animal number=10).

FIG. 6 provides experimental results for the pancreatic tumor growthinhibition studies showing the effects of MNTX1 on AsPC-1 pancreaticcancer. FIG. 6 shows tumor weight for control conditions correspondingto intraperitoneal injection of PBS and conditions for corresponding tointraperitoneal injection of MNTX1. The figure shows the tumor weightmeasured at sacrifice. The treatment with MNTX1 was measurably superiorto the control. FIG. 7 provides AsPC1 pancreatic tumor size photographrecords at the end point of the mouse sacrifice observed for pancreatictumor growth inhibition studies.

Example 5 Administration and Formulation

5a: Salts and Prodrugs

The invention contemplates pharmaceutically active compounds eitherchemically synthesized or formed by in vivo biotransformation tocompounds set forth herein.

Compounds of this invention and compounds useful in the methods of thisinvention include those of the compounds and formula (s) describedherein and pharmaceutically-acceptable salts and esters of thosecompounds. In embodiments, salts include any salts derived from theacids and bases of the formulas herein which are acceptable for use inhuman or veterinary applications. In embodiments, the term ester refersto hydrolyzable esters of compounds of the names and formulas herein. Inembodiments, salts and esters of the compounds of the formulas hereincan include those which have the same or better therapeutic, diagnostic,or pharmaceutical (human or veterinary) general properties as thecompounds of the formulas herein. In an embodiment, a composition of theinvention is a compound or salt or ester thereof suitable forpharmaceutical formulations.

Compounds of the invention can have prodrug forms. Prodrugs of thecompounds of the invention are useful in embodiments includingcompositions and methods. Any compound that will be converted in vivo toprovide a biologically, pharmaceutically, diagnostically, ortherapeutically active form of a compound of the invention is a prodrug.Various examples and forms of prodrugs are well known in the art.Examples of prodrugs are found, inter alia, in: Design of Prodrugs,edited by H. Bundgaard, (Elsevier, 1985); Methods in Enzymology, Vol.42, at pp. 309-396, edited by K. Widder, et. al. (Academic Press, 1985);A Textbook of Drug Design and Development, edited by Krosgaard-Larsenand H. Bundgaard, Chapter 5, “Design and Application of Prodrugs,” by H.Bundgaard, at pp. 113-191 (1991); H. Bundgaard, Advanced Drug DeliveryReviews, Vol. 8, p. 1-38 (1992); H. Bundgaard, et al., Journal ofPharmaceutical Sciences, Vol. 77, p. 285 (1988); and Nogrady (1985)Medicinal Chemistry A Biochemical Approach, Oxford University Press, NewYork, pages 388-392). A prodrug, such as a pharmaceutically acceptableprodrug, can represent prodrugs of the compounds of the invention whichare, within the scope of sound medical judgment, suitable for use incontact with the tissues of humans and lower animals without unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use.Prodrugs of the invention can be rapidly transformed in vivo to a parentcompound of a compound described herein, for example, by hydrolysis inblood or by other cell, tissue, organ, or system processes. Furtherdiscussion is provided in: T. Higuchi and V. Stella, Pro-drugs as NovelDelivery Systems, V. 14 of the A.C.S. Symposium Series; and in Edward B.Roche, ed., Bioreversible Carriers in Drug Design, AmericanPharmaceutical Association and Pergamon Press (1987).

Optical agents of the invention can be formulated withpharmaceutically-acceptable anions and/or cations.Pharmaceutically-acceptable cations include among others, alkali metalcations (e.g., Li⁺, Na⁺, K⁺), alkaline earth metal cations (e.g., Ca²⁺,Mg²⁺), non-toxic heavy metal cations and ammonium (NH₄ ⁺) andsubstituted ammonium (N(R′)₄ ⁺, where R′ is hydrogen, alkyl, orsubstituted alkyl, i.e., including, methyl, ethyl, or hydroxyethyl,specifically, trimethyl ammonium, triethyl ammonium, and triethanolammonium cations). Pharmaceutically-acceptable anions include, amongothers, halides (e.g., F⁻, Cl⁻, Br⁻, At⁻), sulfate, acetates (e.g.,acetate, trifluoroacetate), ascorbates, aspartates, benzoates, citrates,and lactate.

Pharmaceutically acceptable salts comprise pharmaceutically-acceptableanions and/or cations. As used herein, the term “pharmaceuticallyacceptable salt” can refer to acid addition salts or base addition saltsof the compounds in the present disclosure. A pharmaceuticallyacceptable salt is any salt which retains at least a portion of theactivity of the parent compound and does not impart significantdeleterious or undesirable effect on a subject to whom it isadministered and in the context in which it is administered.Pharmaceutically acceptable salts include metal complexes and salts ofboth inorganic and organic acids. Pharmaceutically acceptable saltsinclude metal salts such as aluminum, calcium, iron, magnesium,manganese and complex salts. Pharmaceutically acceptable salts include,but are not limited to, acid salts such as acetic, aspartic,alkylsulfonic, arylsulfonic, axetil, benzenesulfonic, benzoic,bicarbonic, bisulfuric, bitartaric, butyric, calcium edetate, camsylic,carbonic, chlorobenzoic, cilexetil, citric, edetic, edisylic, estolic,esyl, esylic, formic, fumaric, gluceptic, gluconic, glutamic, glycolic,hexamic, hexylreserinoic, hydrobromic, hydrochloric, hydroiodic,hydroxynaphthoic, isethionic, lactic, lactobionic, maleic, malic,malonic, mandelic, methanesulfonic, mucic, muconic, napsylic, nitric,oxalic, p-nitromethanesulfonic, palmoic, pantothenic, phosphoric,monohydrogen phosphoric, dihydrogen phosphoric, phthalic,polygalactouronic, propionic, salicylic, stearic, succinic, sulfamic,sulfanlic, sulfonic, sulfuric, tannic, tartaric, teoclic,toluenesulfonic, polyglutamic, polyaspartic and the like.Pharmaceutically acceptable salts can be derived from amino acids,including, but not limited to, cysteine. Other pharmaceuticallyacceptable salts can be found, for example, in Stahl et al., Handbook ofPharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH, VerlagHelvetica Chimica Acta, Zürich, 2002. (ISBN 3-906390-26-8).

5b: Efficacy

Typically, a compound of the invention, or pharmaceutically acceptablesalt thereof, is administered to a subject in a diagnostically ortherapeutically effective amount. One skilled in the art generally candetermine an appropriate dosage.

Compositions for oral administration can be, for example, prepared in amanner such that a single dose in one or more oral preparations containsat least about 20 mg of the anti-angiogenesis agent compound per squaremeter of subject body surface area, or at least about 50, 100, 150, 200,300, 400, or 500 mg of the anti-angiogenesis agent compound per squaremeter of subject body surface area (the average body surface area for ahuman is, for example, 1.8 square meters). In particular, a single doseof a composition for oral administration can contain from about 20 toabout 600 mg, and in certain aspects from about 20 to about 400 mg, inanother aspect from about 20 to about 300 mg, and in yet another aspectfrom about 20 to about 200 mg of the anti-angiogenesis agent compoundper square meter of subject body surface area. Compositions forparenteral administration can be prepared in a manner such that a singledose contains at least about 20 mg of the anti-angiogenesis agentcompound per square meter of subject body surface area, or at leastabout 40, 50, 100, 150, 200, 300, 400, or 500 mg of theanti-angiogenesis agent compound per square meter of subject bodysurface area. In particular, a single dose in one or more parenteralpreparations contains from about 20 to about 500 mg, and in certainaspects from about 20 to about 400 mg, and in another aspect from about20 to about 450 mg, and in yet another aspect from about 20 to about 350mg of the anti-angiogenesis agent per square meter of subject bodysurface area. It should be recognized that these oral and parenteraldosage ranges represent generally preferred dosage ranges, and are notintended to limit the invention. The dosage regimen actually employedcan vary widely, and, therefore, can deviate from the generallypreferred dosage regimen. It is contemplated that one skilled in the artwill tailor these ranges to the individual subject.

Toxicity and therapeutic efficacy of such compounds and bioconjugatescan be determined by standard pharmaceutical procedures in cell culturesor experimental animals for determining the LD₅₀ (the dose lethal to 50%of the population) and the ED₅₀, (the dose therapeutically effective in50% of the population). The dose ratio between toxic and therapeuticeffects is the therapeutic index that can be expressed as the ratioLD₅₀/ED₅₀. Compounds and bioconjugates that exhibit large therapeuticindices are preferred. While compounds and bioconjugates exhibitingtoxic side effects can be used, care should be taken to design adelivery system that targets such compounds and bioconjugates to thesite affected by the disease or disorder in order to minimize potentialdamage to unaffected cells and reduce side effects.

Data obtained from the cell culture assays and animal studies can beused in formulating a range of dosages for use in humans and othermammals. The dosage of such compounds and bioconjugates lies preferablywithin a range of circulating plasma or other bodily fluidconcentrations that include the ED₅₀ and provides clinically efficaciousresults (i.e., reduction in disease symptoms). The dosage can varywithin this range depending upon the dosage form employed and the routeof administration utilized. For any compound and bioconjugate of thepresent invention, the therapeutically effective amount can be estimatedinitially from cell culture assays. A dosage can be formulated in animalmodels to achieve a circulating plasma concentration range that includesthe ED₅₀ (the concentration of the test compound that achieves ahalf-maximal inhibition of symptoms) as determined in cell culture. Suchinformation can be used to more accurately determine useful dosages inhumans and other mammals. Compound and bioconjugate levels in plasma canbe measured, for example, by high performance liquid chromatography.

An amount of a compound or bioconjugate that can be combined with apharmaceutically acceptable carrier to produce a single dosage form willvary depending upon the patient treated and the particular mode ofadministration. It will be appreciated by those skilled in the art thatthe unit content of a compound/bioconjugate contained in an individualdose of each dosage form need not in itself constitute a therapeuticallyeffective amount, as the necessary therapeutically effective amountcould be reached by administration of a number of individual doses. Theselection of dosage depends upon the dosage form utilized, the conditionbeing treated, and the particular purpose to be achieved according tothe determination of those skilled in the art.

The dosage and dosage regime for treating a disease or condition can beselected in accordance with a variety of factors, including the type,age, weight, sex, diet and/or medical condition of the patient, theroute of administration, pharmacological considerations such asactivity, efficacy, pharmacokinetic and/or toxicology profiles of theparticular compound/bioconjugate employed, whether acompound/bioconjugate delivery system is utilized, and/or whether thecompound/bioconjugate is administered as a pro-drug or part of a drugcombination. Thus, the dosage regime actually employed can vary widelyfrom subject to subject, or disease to disease and different routes ofadministration can be employed in different clinical settings.

The identified compounds/bioconjugates monitor, treat, inhibit, controland/or prevent, or at least partially arrest or partially prevent,diseases and conditions of interest and can be administered to a subjectat therapeutically effective amounts and optionally diagnosticallyeffective amounts. Compositions/formulations of the present inventioncomprise a therapeutically effective amount (which can optionallyinclude a diagnostically effective amount) of at least one compound orbioconjugate of the present invention. Subjects receiving treatment thatincludes a compound/bioconjugate of the invention are preferably animals(e.g., mammals, reptiles and/or avians), more preferably humans, horses,cows, dogs, cats, sheep, pigs, and/or chickens, and most preferablyhumans.

5c: Administration

The preferred composition depends on the route of administration. Anyroute of administration can be used as long as the target of thecompound or pharmaceutically acceptable salt is available via thatroute. Suitable routes of administration include, for example, oral,intravenous, parenteral, inhalation, rectal, nasal, topical (e.g.,transdermal and intraocular), intravesical, intrathecal, enteral,pulmonary, intralymphatic, intracavital, vaginal, transurethral,intradermal, aural, intramammary, buccal, orthotopic, intratracheal,intralesional, percutaneous, endoscopical, transmucosal, sublingual, andintestinal administration.

In an embodiment, the invention provides a method for treating a medicalcondition comprising administering to a subject (e.g. patient) in needthereof, a therapeutically effective amount of a composition of theinvention, such as a compound of any one of formulas (FX1)-(FX164). Inan embodiment, the invention provides a method for diagnosing or aidingin the diagnosis of a medical condition comprising administering to asubject in need thereof, a diagnostically effective amount of acomposition of the invention. In an embodiment, the medical condition iscancer, or various other diseases, injuries, and disorders, includingcardiovascular disorders such as atherosclerosis and vascularrestenosis, inflammatory diseases, ophthalmic diseases anddermatological diseases.

The diagnostic and therapeutic formulations of this invention can beadministered alone, but can be administered with a pharmaceuticalcarrier selected upon the basis of the chosen route of administrationand standard pharmaceutical practice.

Any suitable form of administration can be employed in connection withthe diagnostic and therapeutic formulations of the invention. Thediagnostic and therapeutic formulations of this invention can beadministered intravenously, in oral dosage forms, intraperitoneally,subcutaneously, or intramuscularly, all using dosage forms well known tothose of ordinary skill in the pharmaceutical arts.

The present compositions, preparations and formulations can beformulated into diagnostic or therapeutic compositions for enteral,parenteral, topical, aerosol, inhalation, or cutaneous administration.Topical or cutaneous delivery of the compositions, preparations andformulations can also include aerosol formulation, creams, gels,solutions, etc. The present compositions, preparations and formulationsare administered in doses effective to achieve the desired diagnosticand/or therapeutic effect. Such doses can vary widely depending upon theparticular compositions employed in the composition, the organs ortissues to be examined, the equipment employed in the clinicalprocedure, the efficacy of the treatment achieved, and the like. Thesecompositions, preparations and formulations contain an effective amountof the composition(s), along with conventional pharmaceutical carriersand excipients appropriate for the type of administration contemplated.These compositions, preparations and formulations can also optionallyinclude stabilizing agents and skin penetration enhancing agents.

(i) Parenteral Administration

Compounds and bioconjugates of the present invention can be formulatedfor parenteral administration by injection (e.g., by bolus injection orcontinuous infusion). Formulations for injection can be presented inunit dosage form in ampoules or in multi-dose containers with anoptional preservative added. The parenteral preparation can be enclosedin ampoules, disposable syringes or multiple dose vials made of glass,plastic or the like. The formulation can take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and can containformulatory agents such as suspending, stabilizing and/or dispersingagents.

For example, a parenteral preparation can be a sterile injectablesolution or suspension in a nontoxic parenterally acceptable diluent orsolvent (e.g., as a solution in 1,3-butanediol). Among the acceptablevehicles and solvents that can be employed are water, Ringer's solution,and isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil can be employed including synthetic mono- ordi-glycerides. In addition, fatty acids such as oleic acid can be usedin the parenteral preparation.

Alternatively, compounds and bioconjugates of the present invention canbe formulated in powder form for constitution with a suitable vehicle,such as sterile pyrogen-free water, before use. For example, acompound/bioconjugate suitable for parenteral administration can includea sterile isotonic saline solution containing between 0.1 percent and 90percent weight per volume of the compound/bioconjugate. By way ofexample, a solution can contain from about 5 percent to about 20percent, more preferably from about 5 percent to about 17 percent, morepreferably from about 8 to about 14 percent, and still more preferablyabout 10 percent weight per volume of the compound/bioconjugate. Thesolution or powder preparation can also include a solubilizing agent anda local anesthetic such as lignocaine to ease pain at the site of theinjection. Other methods of parenteral delivery ofcompounds/bioconjugates will be known to the skilled artisan and arewithin the scope of the invention.

(ii) Oral Administration

For oral administration, a compound/bioconjugate of the invention can beformulated to take the form of tablets or capsules prepared byconventional means with one or more pharmaceutically acceptable carriers(e.g., excipients such as binding agents, fillers, lubricants anddisintegrants).

(iii) Controlled-Release Administration

Controlled-release (or sustained-release) preparations can be formulatedto extend the activity of a compound/bioconjugate and reduce dosagefrequency. Controlled-release preparations can also be used to effectthe time of onset of action or other characteristics, such as bloodlevels of the compound/bioconjugate, and consequently affect theoccurrence of side effects.

Controlled-release preparations can be designed to initially release anamount of a compound/bioconjugate that produces the desired therapeuticeffect, and gradually and continually release other amounts of thecompound/bioconjugate to maintain the level of therapeutic effect overan extended period of time. In order to maintain a near-constant levelof a compound/bioconjugate in the body, the compound/bioconjugate can bereleased from the dosage form at a rate that will replace the amount ofcompound/bioconjugate being metabolized and/or excreted from the body.The controlled-release of a compound/bioconjugate can be stimulated byvarious inducers, e.g., change in pH, change in temperature, enzymes,water, and/or other physiological conditions or molecules.

Controlled-release systems can include, for example, an infusion pumpwhich can be used to administer the compound/bioconjugate in a mannersimilar to that used for delivering insulin or chemotherapy to the bodygenerally, or to specific organs or tumors. Typically, using such asystem, the compound/bioconjugate is administered in combination with abiodegradable, biocompatible polymeric implant that releases thecompound/bioconjugate over a controlled period of time at a selectedsite. Examples of polymeric materials include polyanhydrides,polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinylacetate, (PEG) polyethyleneglycol and copolymers and combinationsthereof. In addition, a controlled release system can be placed inproximity of a therapeutic target (e.g., organ, tissue, or group ofcells), thus requiring only a fraction of a systemic dosage.

Compounds/bioconjugates of the invention can be administered by othercontrolled-release means or delivery devices that are well known tothose of ordinary skill in the art. These include, for example,hydropropylmethyl cellulose, other polymer matrices, gels, permeablemembranes, osmotic systems, multilayer coatings, microparticles,liposomes, microspheres, or the like, or a combination of any of theabove to provide the desired release profile in varying proportions.Other methods of controlled-release delivery of compounds/bioconjugateswill be known to the skilled artisan and are within the scope of theinvention.

(iv) Inhalation Administration

Compounds/bioconjugates of the invention can be administered directly tothe lung of a patient/subject by inhalation. For administration byinhalation, a compound/bioconjugate can be conveniently delivered to thelung by a number of different devices. For example, a Metered DoseInhaler (“MDI”) which utilizes canisters that contain a suitable lowboiling point propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas can be used to deliver a compound/bioconjugatedirectly to the lung. MDI devices are available from a number ofsuppliers such as 3M Corporation, Aventis, Boehringer Ingleheim, ForestLaboratories, GlaxoSmithKline, Merck & Co. and Vectura.

Alternatively, a Dry Powder Inhaler (DPI) device can be used toadminister a compound/bioconjugate to the lung. DPI devices typicallyuse a mechanism such as a burst of gas to create a cloud of dry powderinside a container, which can then be inhaled by the patient. DPIdevices are also well known in the art and can be purchased from anumber of vendors which include, for example, GlaxoSmithKline, NektarTherapeutics, Innovata and Vectura. A popular variation is the multipledose DPI (“MDDPI”) system, which allows for the delivery of more thanone therapeutic dose. MDDPI devices are available from companies such asAstraZeneca, GlaxoSmithKline, TEVA, Merck & Co., SkyePharma and Vectura.For example, capsules and cartridges of gelatin for use in an inhaler orinsufflator can be formulated containing a powder mix of thecompound/bioconjugate and a suitable powder base such as lactose orstarch for these systems.

Another type of device that can be used to deliver acompound/bioconjugate to the lung is a liquid spray device supplied, forexample, by Aradigm Corporation. Liquid spray systems use extremelysmall nozzle holes to aerosolize liquid compound/bioconjugateformulations that can then be directly inhaled into the lung. Forexample, a nebulizer device can be used to deliver acompound/bioconjugate to the lung. Nebulizers create aerosols fromliquid compound/bioconjugate formulations by using, for example,ultrasonic energy to form fine particles that can be readily inhaled.Examples of nebulizers include devices supplied by Aventis and Battelle.

In another example, an electrohydrodynamic (“EHD”) aerosol device can beused to deliver a compound/bioconjugate to the lung. END aerosol devicesuse electrical energy to aerosolize liquid compound/bioconjugatesolutions or suspensions. The electrochemical properties of thecompound/bioconjugate formulation are important parameters to optimizewhen delivering this compound/bioconjugate to the lung with an EHDaerosol device. Such optimization is routinely performed by one of skillin the art. Other methods of intra-pulmonary delivery ofcompounds/bioconjugates will be known to the skilled artisan and arewithin the scope of the invention.

Liquid compound/bioconjugate formulations suitable for use withnebulizers and liquid spray devices and EHD aerosol devices willtypically include the compound/bioconjugate with a pharmaceuticallyacceptable carrier. In one exemplary embodiment, the pharmaceuticallyacceptable carrier is a liquid such as alcohol, water, polyethyleneglycol or a perfluorocarbon. Optionally, another material can be addedto alter the aerosol properties of the solution or suspension of thecompound/bioconjugate. For example, this material can be a liquid suchas an alcohol, glycol, polyglycol or a fatty acid. Other methods offormulating liquid compound/bioconjugate solutions or suspensionssuitable for use in aerosol devices are known to those of skill in theart.

(v) Depot Administration

A compound/bioconjugate of the invention can be formulated as a depotpreparation. Such long-acting formulations can be administered byimplantation (e.g., subcutaneously or intramuscularly) or byintramuscular injection. Accordingly, the compound/bioconjugate can beformulated with suitable polymeric or hydrophobic materials such as anemulsion in an acceptable oil or ion exchange resin, or as sparinglysoluble derivatives such as a sparingly soluble salt. Other methods ofdepot delivery of compounds/bioconjugates will be known to the skilledartisan and are within the scope of the invention.

(vi) Topical Administration

For topical application, a compound/bioconjugate can be combined with apharmaceutically acceptable carrier so that an effective dosage isdelivered, based on the desired activity ranging from an effectivedosage, for example, of 1.0 μM to 1.0 mM. In one aspect of theinvention, a topical formulation of a compound/bioconjugate can beapplied to the skin. The pharmaceutically acceptable carrier can be inthe form of, for example, and not by way of limitation, an ointment,cream, gel, paste, foam, aerosol, suppository, pad or gelled stick.

A topical formulation can include a therapeutically effective amount ofa compound/bioconjugate in an ophthalmologically acceptable excipientsuch as buffered saline, mineral oil, vegetable oils such as corn orarachis oil, petroleum jelly, Miglyol 182, alcohol solutions, orliposomes or liposome-like products. Any of these formulations of suchcompounds/bioconjugates can include preservatives, antioxidants,antibiotics, immunosuppressants, and other biologically orpharmaceutically effective agents that do not exert a significantdetrimental effect on the compound/bioconjugate. Other methods oftopical delivery of compounds/bioconjugates will be known to the skilledartisan and are within the scope of the invention.

(vii) Rectal Administration

Compounds/bioconjugates of the invention can be formulated in rectalformulations such as suppositories or retention enemas that includeconventional suppository bases such as cocoa butter or other glyceridesand/or binders and/or carriers such as triglycerides, microcrystallinecellulose, gum tragacanth or gelatin. Rectal formulations can contain acompound/bioconjugate in the range of 0.5% to 10% by weight, forexample. Other methods of rectal delivery of compounds/bioconjugateswill be known to the skilled artisan and are within the scope of theinvention.

(viii) Other Systems of Administration

Various other delivery systems are known in the art and can be used toadminister the compounds/bioconjugates of the invention. Moreover, theseand other delivery systems can be combined and/or modified to promoteoptimization of the administration of compounds/bioconjugates of thepresent invention. Exemplary formulations that includecompounds/bioconjugates of the present invention are described elsewhereherein (the compounds/bioconjugates of the present invention areindicated as the active ingredient, but those of skill in the art willrecognize that pro-drugs and compound combinations are also meant to beencompassed by this term).

5d: Formulation

In an embodiment, the invention provides a medicament which comprises atherapeutically effective amount of one or more compositions of theinvention, such as a compound of any one of formulas (FX1)-(FX164). Inan embodiment, the invention provides a medicament which comprises adiagnostically effective amount of one or more compositions of theinvention. In an embodiment, the invention provides a method for makinga medicament for treatment of a condition described herein. In anembodiment, the invention provides a method for making a medicament fordiagnosis or aiding in the diagnosis of a condition described herein. Inan embodiment, the invention provides the use of one or morecompositions set forth herein for the making of a medicament. In anembodiment, the invention provides the use of one or more compositionsset forth herein for the treatment of a disease. In an embodiment, theinvention provides the use of one or more compositions set forth hereinfor the diagnosis of a disease. Compositions of the invention includeformulations and preparations comprising one or more of the presentoptical agents provided in an aqueous solution, such as apharmaceutically acceptable formulation or preparation. Optionally,compositions of the invention further comprise one or morepharmaceutically acceptable surfactants, buffers, electrolytes, salts,carriers, binders, coatings, preservatives and/or excipients.

In an embodiment, the invention provides a pharmaceutical formulationhaving an active ingredient comprising a composition of the invention,such as a compound of any one of formulas (FX1)-(FX164). In anembodiment, the invention provides a method of synthesizing acomposition of the invention or a pharmaceutical formulation thereof,such as a compound of any one of formulas (FX1)-(FX164). In anembodiment, a pharmaceutical formulation comprises one or moreexcipients, carriers, diluents, and/or other components as would beunderstood in the art. Preferably, the components meet the standards ofthe National Formulary (“NF”), United States Pharmacopoeia (“USP”;United States Pharmacopeial Convention Inc., Rockville, Md.), orHandbook of Pharmaceutical Manufacturing Formulations (Sarfaraz K.Niazi, all volumes, ISBN: 9780849317521, ISBN 10: 0849317525; CRC Press,2004). See, e.g., United States Pharmacopeia and National Formulary (USP30-NF 25), Rockville, Md.: United States Pharmacopeial Convention (2007and 2008), and each of any earlier editions; The Handbook ofPharmaceutical Excipients, published jointly by the American PharmacistsAssociation and the Pharmaceutical Press (Pharmaceutical Press (2005)(ISBN-10: 0853696187, ISBN-13: 978-0853696186)); Merck Index, Merck &Co., Rahway, N.J.; and Gilman et al., (eds) (1996); Goodman andGilman's: The Pharmacological Bases of Therapeutics, 8th Ed., PergamonPress. In embodiments, the formulation base of the formulations of theinvention comprises physiologically acceptable excipients, namely, atleast one binder and optionally other physiologically acceptableexcipients. Physiologically acceptable excipients are those known to beusable in the pharmaceutical technology sectors and adjacent areas,particularly, those listed in relevant pharmacopeias (e.g. DAB, Ph.Eur., BP, NF, USP), as well as other excipients whose properties do notimpair a physiological use.

This invention also is directed, in part, to pharmaceutical compositionsincluding a therapeutically effective amount of a compound or salt ofthis invention, as well as processes for making such compositions. Suchcompositions generally include one or more pharmaceutically acceptablecarriers (e.g., excipients, vehicles, auxiliaries, adjuvants, diluents)and can include other active ingredients. Formulation of thesecompositions can be achieved by various methods known in the art. Ageneral discussion of these methods can be found in, for example,Hoover, John E., Remington's Pharmaceutical Sciences (Mack PublishingCo., Easton, Pa.: 1975). See also, Lachman, L., eds., PharmaceuticalDosage Forms (Marcel Decker, New York, N.Y., 1980).

The diagnostic and therapeutic formulations of this invention andmedicaments of this invention can further comprise one or morepharmaceutically acceptable carriers, excipients, buffers, emulsifiers,surfactants, electrolytes or diluents. Such compositions and medicamentsare prepared in accordance with acceptable pharmaceutical procedures,such as, for example, those described in Remingtons PharmaceuticalSciences, 17th edition, ed. Alfonoso R. Gennaro, Mack PublishingCompany, Easton, Pa. (1985).

Compositions of the invention include formulations and preparationscomprising one or more of the present compounds provided in an aqueoussolution, such as a pharmaceutically acceptable formulation orpreparation. Optionally, compositions of the invention further compriseone or more pharmaceutically acceptable surfactants, buffers,electrolytes, salts, carriers, binders, coatings, preservatives and/orexcipients.

Compounds and bioconjugates of the present invention can be formulatedby known methods for administration to a subject using several routeswhich include, but are not limited to, parenteral, oral, topical,intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous,intranasal, epidural, and ophthalmic routes. An individualcompound/bioconjugate can be administered in combination with one ormore additional compounds/bioconjugates of the present invention and/ortogether with other biologically active or biologically inert agents.Such biologically active or inert agents can be in fluid or mechanicalcommunication with the compound(s)/bioconjugate(s) or attached to thecompound(s)/bioconjugate(s) by ionic, covalent, Van der Waals,hydrophobic, hydrophilic or other physical forces. It is preferred thatadministration is localized in a subject, but administration can also besystemic.

Compounds and bioconjugates of the present invention can be formulatedby any conventional manner using one or more pharmaceutically acceptablecarriers. Thus, the compound(s)/bioconjugate(s) and theirpharmaceutically acceptable salts and solvates can be specificallyformulated for administration, e.g., by inhalation or insufflation(either through the mouth or the nose) or oral, buccal, parenteral orrectal administration. The compounds/bioconjugates can take the form ofcharged, neutral and/or other pharmaceutically acceptable salt forms.Examples of pharmaceutically acceptable carriers include, but are notlimited to, those described in REMINGTON'S PHARMACEUTICAL SCIENCES (A.R. Gennaro, Ed.), 20th edition, Williams & Wilkins Pa., USA (2000).

Compounds and bioconjugates of the present invention can be formulatedin the form of solutions, suspensions, emulsions, tablets, pills,capsules, powders, controlled- or sustained-release formulations and thelike. Such formulations will contain a therapeutically effective amountof the compound/bioconjugate, preferably in purified form, together witha suitable amount of carrier so as to provide the form for properadministration to the patient. The formulation should suit the mode ofadministration.

Pharmaceutically acceptable carriers that can be used in conjunctionwith the compounds of the invention are well known to those of ordinaryskill in the art. Carriers can be selected based on a number of factorsincluding, for example, the particular anti-angiogenesis agentcompound(s) or pharmaceutically acceptable salt(s) used; the compound'sconcentration, stability, and intended bioavailability; the conditionbeing treated; the subject's age, size, and general condition; the routeof administration; etc. A general discussion related to carriers can befound in, for example, J. G. Nairn, Remington's Pharmaceutical Science,pp. 1492-1517 (A. Gennaro, ed., Mack Publishing Co., Easton, Pa.(1985)).

Solid dosage forms for oral administration include, for example,capsules, tablets, gelcaps, pills, dragees, troches, powders, granules,and lozenges. In such solid dosage forms, the compounds orpharmaceutically acceptable salts thereof can be combined with one ormore pharmaceutically acceptable carriers. The compounds andpharmaceutically acceptable salts thereof can be mixed with carriersincluding, but not limited to, lactose, sucrose, starch powder, cornstarch, potato starch, magnesium carbonate, microcrystalline cellulose,cellulose esters of alkanoic acids, cellulose alkyl esters, talc,stearic acid, magnesium stearate, magnesium oxide, sodium and calciumsalts of phosphoric and sulfuric acids, sodium carbonate, agar,mannitol, sorbitol, sodium saccharin, gelatin, acacia gum, alginic acid,sodium alginate, tragacanth, colloidal silicon dioxide, croscarmellosesodium, polyvinylpyrrolidone, and/or polyvinyl alcohol, and thentableted or encapsulated for convenient administration. Such capsules ortablets can contain a controlled-release formulation, as can be providedin a dispersion of the compound or salt in hydroxypropylmethylcellulose. In the case of capsules, tablets, and pills, the dosage formsalso can include buffering agents, such as sodium citrate, or magnesiumor calcium carbonate or bicarbonate. Tablets and pills additionally can,for example, include a coating (e.g., an enteric coating) to delaydisintegration and absorption. The concentration of theanti-angiogenesis agent compound in a solid oral dosage form can be fromabout 5 to about 50% for example, and in certain aspects from about 8 toabout 40%, and in another aspect from about 10 to about 30% by weightbased on the total weight of the composition.

Liquid dosage forms of the compounds of the invention for oraladministration include, for example, pharmaceutically acceptableemulsions, solutions, suspensions, syrups, and elixirs containing inertdiluents commonly used in the art (e.g., water). Such compositions alsocan include adjuvants, such as wetting, emulsifying, suspending,flavoring (e.g., sweetening), and/or perfuming agents. The concentrationof the anti-angiogenesis agent compound in the liquid dosage form can befrom about 0.01 to about 5 mg, and in certain aspects from about 0.01 toabout 1 mg, and in another aspect from about 0.01 to about 0.5 mg per mlof the composition. Low concentrations of the compounds of the inventionin liquid dosage form can be prepared in the case that theanti-angiogenesis agent compound is more soluble at low concentrations.Techniques for making oral dosage forms useful in the invention aregenerally described in, for example, Modern Pharmaceutics, Chapters 9and 10 (Banker & Rhodes, Editors (1979)). See also, Lieberman et al.,Pharmaceutical Dosage Forms: Tablets (1981). See also, Ansel,Introduction to Pharmaceutical Dosage Forms (2nd Edition (1976)).

In some aspects of the invention, tablets or powders for oraladministration can be prepared by dissolving the anti-angiogenesis agentcompound in a pharmaceutically acceptable solvent capable of dissolvingthe compound to form a solution and then evaporating when the solutionis dried under vacuum. A carrier can also be added to the solutionbefore drying. The resulting solution can be dried under vacuum to forma glass. The glass can then be mixed with a binder to form a powder.This powder can be mixed with fillers or other conventional tabletingagents, and then processed to form a tablet. Alternatively, the powdercan be added to a liquid carrier to form a solution, emulsion,suspension, or the like.

In some aspects, solutions for oral administration are prepared bydissolving the anti-angiogenesis agent compound in a pharmaceuticallyacceptable solvent capable of dissolving the compound to form asolution. An appropriate volume of a carrier is added to the solutionwhile stirring to form a pharmaceutically acceptable solution for oraladministration.

In some embodiments, a liposome or micelle can be utilized as a carrieror vehicle for the composition. For example, in some embodiments, theanti-angiogenesis agent compound can be a part of the lipophilicbilayers or micelle, and the targeting ligand, if present, can be on theexternal surface of the liposome or micelle. As another example, atargeting ligand can be externally attached to the liposome or micelleafter formulation for targeting the liposome or micelle (which containsthe anti-angiogenesis agent optical agents) to the desired tissue,organ, or other site in the body.

Injectable preparations (e.g., sterile injectable aqueous or oleaginoussuspensions) can be formulated according to the known art using suitabledispersing, wetting agents, and/or suspending agents. Acceptablevehicles for parenteral use include both aqueous and nonaqueouspharmaceutically-acceptable solvents. Suitable pharmaceuticallyacceptable aqueous solvents include, for example, water, salinesolutions, dextrose solutions (such as DW5), electrolyte solutions, etc.

In one embodiment, the present anti-angiogenesis agent compounds areformulated as nanoparticles or microparticles. Use of such nanoparticleor microparticle formulations can be beneficial for some applications toenhance delivery, localization, target specificity, administration, etc.of the anti-angiogenesis agent compound. Potentially usefulnanoparticles and microparticles include, but are not limited to,micelles, liposomes, microemulsions, nanoemulsions, vesicles, tubularmicelles, cylindrical micelles, bilayers, folded sheets structures,globular aggregates, swollen micelles, inclusion complex, encapsulateddroplets, microcapsules, nanocapsules or the like. As will be understoodby those having skill in the art, the present anti-angiogenesis agentcompounds can be located inside the nanoparticle or microparticle,within a membrane or wall of the nanoparticle or microparticle, oroutside of (but bonded to or otherwise associated with) the nanoparticleor microparticle. The agent formulated in nanoparticles ormicroparticles can be administered by any of the routes previouslydescribed. In a formulation applied topically, the anti-angiogenesisagent compound is slowly released over time. In an injectableformulation, the liposome, micelle, capsule, etc., circulates in thebloodstream and is delivered to the desired site (e.g., target tissue).

Preparation and loading of nanoparticles and microparticles are wellknown in the art. As one example, liposomes can be prepared fromdipalmitoyl phosphatidylcholine (DPPC) or egg phosphatidylcholine (PC)because this lipid has a low heat transition. Liposomes are made usingstandard procedures as known to one skilled in the art (e.g.,Braun-Falco et al., (Eds.), Griesbach Conference, Liposome Dermatics,Springer-Verlag, Berlin (1992), pp. 69 81; 91 117. Polycaprolactone,poly(glycolic) acid, poly(lactic) acid, polyanhydride or lipids can beformulated as microspheres. As an illustrative example, the presentanti-angiogenesis agent compounds can be mixed with polyvinyl alcohol(PVA), the mixture then dried and coated with ethylene vinyl acetate,then cooled again with PVA. In a liposome, the present anti-angiogenesisagent compounds can be within one or both lipid bilayers, in the aqueousbetween the bilayers, or within the center or core. Liposomes can bemodified with other molecules and lipids to form a cationic liposome.Liposomes can also be modified with lipids to render their surface morehydrophilic which increases their circulation time in the bloodstream.The thus-modified liposome has been termed a “stealth” liposome, or along-lived liposome, as described in U.S. Pat. No. 6,258,378, and inStealth Liposomes, Lasic and Martin (Eds.) 1995 CRC Press, London.Encapsulation methods include detergent dialysis, freeze drying, filmforming, injection, as known to one skilled in the art and disclosed in,for example, U.S. Pat. No. 6,406,713. Optionally, the presentcompositions and methods include a micelle delivery system, for example,involving one or more PEG-based amphiphilic polymers developed for drugdelivery including: PEG-poly(ε-caprolactone), PEG-poly(amino acid),PEG-polylactide or PEG-phospholipid constructs; a cross linkedpoly(acrylic acid) polymer system, a phospholipid-based system and/orblock copolymer systems comprising one or more of the following polymerblocks: a poly(lactic acid) polymer block; a poly(propylene glycol)polymer block; a poly(amino acid) polymer block; a poly(ester) polymerblock; a poly(ε-caprolactone) polymer block; a poly(ethylene glycol)block, a poly(acrylic acid) block; a polylactide block; a polyesterblock; a polyamide block; a polyanhydride block; a polyurethane block; apolyimine block; a polyurea block; a polyacetal block; a polysaccharideblock; and a polysiloxane block.

Suitable pharmaceutically-acceptable nonaqueous solvents include, butare not limited to, the following (as well as mixtures thereof):

(i) Alcohols (these include, for example, σ-glycerol formal, β-glycerolformal, 1,3-butyleneglycol, aliphatic or aromatic alcohols having from 2to about 30 carbons (e.g., methanol, ethanol, propanol, isopropanol,butanol, t-butanol, hexanol, octanol, amylene hydrate, benzyl alcohol,glycerin (glycerol), glycol, hexylene, glycol, tetrahydrofuranylalcohol, cetyl alcohol, and stearyl alcohol), fatty acid esters of fattyalcohols (e.g., polyalkylene glycols, such as polypropylene glycol andpolyethylene glycol), sorbitan, sucrose, and cholesterol);

(ii) Amides, which include, for example, dimethylacetamide (DMA), benzylbenzoate DMA, dimethylformamide, N-hydroxyethyO-lactamide,N,N-dimethylacetamide-amides, 2-pyrrolidinone, 1-methyl-2-pyrrolidinone,and polyvinylpyrrolidone;

(iii) Esters, which include, for example, acetate esters (e.g.,monoacetin, diacetin, and triacetin), aliphatic and aromatic esters(e.g., ethyl caprylate or octanoate, alkyl oleate, benzyl benzoate, orbenzyl acetate), dimethylsulfoxide (DMSO), esters of glycerin (e.g.,mono, di, and tri-glyceryl citrates and tartrates), ethyl benzoate,ethyl acetate, ethyl carbonate, ethyl lactate, ethyl oleate, fatty acidesters of sorbitan, glyceryl monostearate, glyceride esters (e.g., mono,di, or tri-glycerides), fatty acid esters (e.g., isopropyl myristrate),fatty acid derived PEG esters (e.g., PEG-hydroxyoleate andPEG-hydroxystearate), N-methylpyrrolidinone, pluronic 60,polyoxyethylene sorbitol oleic polyesters (e.g., poly(ethoxylated)₃₀₋₆₀sorbitol poly(oleate)₂₋₄, poly(oxyethylene)₁₅₋₂₀ monooleate,poly(oxyethylene)₁₅₋₂₀ mono 12-hydroxystearate, andpoly(oxyethylene)₁₅₋₂₀ mono ricinoleate), polyoxyethylene sorbitanesters (e.g., polyoxyethylene-sorbitan monooleate,polyoxyethylene-sorbitan monopalmitate, polyoxyethylene-sorbitanmonolaurate, polyoxyethylene-sorbitan monostearate, and POLYSORBATE 20,40, 60, and 80 (from ICI Americas, Wilmington, Del.)),polyvinylpyrrolidone, alkyleneoxy modified fatty acid esters (e.g.,polyoxyl 40 hydrogenated castor oil and polyoxyethylated castor oils,such as CREMOPHOR EL solution or CREMOPHOR RH 40 solution), saccharidefatty acid esters (i.e., the condensation product of a monosaccharide(e.g., pentoses, such as, ribose, ribulose, arabinose, xylose, lyxose,and xylulose; hexoses, such as glucose, fructose, galactose, mannose,and sorbose; trioses; tetroses; heptoses; and octoses), disaccharide(e.g., sucrose, maltose, lactose, and trehalose), oligosaccharide, or amixture thereof with one or more C₄-C₂₂ fatty acids (e.g., saturatedfatty acids, such as caprylic acid, capric acid, lauric acid, myristicacid, palmitic acid, and stearic acid; and unsaturated fatty acids, suchas palmitoleic acid, oleic acid, elaidic acid, erucic acid, and linoleicacid), and steroidal esters;

(iv) Ethers, for example, alkyl, aryl, and cyclic ethers having from 2to about 30 carbons. Examples include diethyl ether, tetrahydrofuran,dimethyl isosorbide, diethylene glycol monoethyl ether), and glycofurol(tetrahydrofurfuranyl alcohol polyethylene glycol ether);

(v) Ketones which typically have from about 3 to about 30 carbons.Examples include acetone, methyl ethyl ketone, and methyl isobutylketone;

(vi) Hydrocarbons which are typically aliphatic, cycloaliphatic, oraromatic hydrocarbons having from about 4 to about 30 carbons. Examplesinclude benzene, cyclohexane, dichloromethane, dioxolanes, hexane,n-decane, n-dodecane, n-hexane, sulfolane, tetramethylenesulfone,tetramethylenesulfoxide, toluene, dimethylsulfoxide (DMSO); andtetramethylene sulfoxide;

(vii) Oils which include, for example, oils of mineral, vegetable,animal, essential, or synthetic origin. These include: mineral oils,such as aliphatic and wax-based hydrocarbons, aromatic hydrocarbons,mixed aliphatic and aromatic based hydrocarbons, and refined paraffinoil; vegetable oils, such as linseed, tung, safflower, soybean, castor,cottonseed, groundnut, rapeseed, coconut, palm, olive, corn, corn germ,sesame, persic, and peanut oil; glycerides, such as mono-, di-, andtriglycerides; animal oils, such as fish, marine, sperm, cod-liver,haliver, squalene, squalane, and shark liver oil; oleic oils; andpolyoxyethylated castor oil;

(viii) Alkyl, alkenyl, or aryl halides which include, for example, alkylor aryl halides having from 1 to about 30 carbons and one or morehalogen substituents. Examples include: methylene chloride;monoethanolamine; petroleum benzin; trolamine; omega-3 polyunsaturatedfatty acids (e.g., alpha-linolenic acid, eicosapentaenoic acid,docosapentaenoic acid, or docosahexaenoic acid); polyglycol ester of12-hydroxystearic acid and polyethylene glycol (SOLUTOL HS-15, fromBASF, Ludwigshafen, Germany); polyoxyethylene glycerol; sodium laurate;sodium oleate; and sorbitan monooleate.

Other pharmaceutically acceptable solvents for use in the invention arewell known to those of ordinary skill in the art. General discussionrelating to such solvents can be found in, for example, The ChemotherapySource Book (Williams & Wilkens Publishing), The Handbook ofPharmaceutical Excipients, (American Pharmaceutical Association,Washington, D.C., and The Pharmaceutical Society of Great Britain,London, England, 1968), Modern Pharmaceutics 3d ed., (G. Banker et. al.,eds., Marcel Dekker, Inc., New York, N.Y. (1995)), The PharmacologicalBasis of Therapeutics, (Goodman & Gilman, McGraw Hill Publishing),Pharmaceutical Dosage Forms, (H. Lieberman et. al., eds., Marcel Dekker,Inc., New York, N.Y. (1980)), Remington's Pharmaceutical Sciences, 19thed., (A. Gennaro, ed., Mack Publishing, Easton, Pa., (1995)), The UnitedStates Pharmacopeia 24, The National Formulary 19, (National Publishing,Philadelphia, Pa. (2000)); Spiegel, A. J., et al., “Use of NonaqueousSolvents in Parenteral Products,” J. Pharma. Sciences, Vol. 52, No. 10,pp. 917-927 (1963).

Solvents useful in the invention include, but are not limited to, thoseknown to stabilize anti-angiogenesis agent compounds or pharmaceuticallyacceptable salts thereof. These can include, for example, oils rich intriglycerides, such as safflower oil, soybean oil, and mixtures thereof;and alkyleneoxy-modified fatty acid esters, such as polyoxyl 40hydrogenated castor oil and polyoxyethylated castor oils (e.g.,CREMOPHOR EL solution or CREMOPHOR RH 40 solution). Commerciallyavailable triglycerides include INTRALIPID emulsified soybean oil(Kabi-Pharmacia Inc., Stockholm, Sweden), NUTRALIPID emulsion (McGaw,Irvine, Calif.), LIPOSYN II 20% emulsion (a 20% fat emulsion solutioncontaining 100 mg safflower oil, 100 mg soybean oil, 12 mg eggphosphatides, and 25 mg glycerin per ml of solution; AbbottLaboratories, Chicago, Ill.), LIPOSYN III 2% emulsion (a 2% fat emulsionsolution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg eggphosphatides, and 25 mg glycerin per ml of solution; AbbottLaboratories, Chicago, Ill.), natural or synthetic glycerol derivativescontaining the docosahexaenoyl group at levels of from about 25 to about100% (by weight based on the total fatty acid content) (DHASCO fromMartek Biosciences Corp., Columbia, Md.; DHA MAGURO from DaitoEnterprises, Los Angeles, Calif.; SOYACAL; and TRAVEMULSION). Ethanol inparticular is a useful solvent for dissolving a anti-angiogenesis agentcompound or pharmaceutically acceptable salt thereof to form solutions,emulsions, and the like.

Additional components can be included in the compositions of thisinvention for various purposes generally known in the pharmaceuticalindustry. These components tend to impart properties that, for example,enhance retention of the anti-angiogenesis agent compound or salt at thesite of administration, protect the stability of the composition,control the pH, and facilitate processing of the anti-angiogenesis agentcompound or salt into pharmaceutical formulations, and the like.Specific examples of such components include cryoprotective agents;agents for preventing reprecipitation of the anti-angiogenesis agentcompound or salt surface; active, wetting, or emulsifying agents (e.g.,lecithin, polysorbate-80, TWEEN 80, pluronic 60, and polyoxyethylenestearate); preservatives (e.g., ethyl-p-hydroxybenzoate); microbialpreservatives (e.g., benzyl alcohol, phenol, m-cresol, chlorobutanol,sorbic acid, thimerosal, and paraben); agents for adjusting pH orbuffering agents (e.g., acids, bases, sodium acetate, sorbitanmonolaurate, etc.); agents for adjusting osmolarity (e.g., glycerin);thickeners (e.g., aluminum monostearate, stearic acid, cetyl alcohol,stearyl alcohol, guar gum, methyl cellulose, hydroxypropylcellulose,tristearin, cetyl wax esters, polyethylene glycol, etc.); colorants;dyes; flow aids; non-volatile silicones (e.g., cyclomethicone); clays(e.g., bentonites); adhesives; bulking agents; flavorings; sweeteners;adsorbents; fillers (e.g., sugars such as lactose, sucrose, mannitol,sorbitol, cellulose, calcium phosphate, etc.); diluents (e.g., water,saline, electrolyte solutions, etc.); binders (e.g., gelatin; gumtragacanth; methyl cellulose; hydroxypropyl methylcellulose; sodiumcarboxymethyl cellulose; polyvinylpyrrolidone; sugars; polymers; acacia;starches, such as maize starch, wheat starch, rice starch, and potatostarch; etc.); disintegrating agents (e.g., starches, such as maizestarch, wheat starch, rice starch, potato starch, and carboxymethylstarch; cross-linked polyvinyl pyrrolidone; agar; alginic acid or a saltthereof, such as sodium alginate; croscarmellose sodium; crospovidone;etc); lubricants (e.g., silica; talc; stearic acid and salts thereof,such as magnesium stearate; polyethylene glycol; etc.); coating agents(e.g., concentrated sugar solutions including gum arabic, talc,polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titaniumdioxide, etc.); and antioxidants (e.g., sodium metabisulfite, sodiumbisulfite, sodium sulfite, dextrose, phenols, thiophenols, etc.).

Techniques and compositions for making parenteral dosage forms aregenerally known in the art. Formulations for parenteral administrationcan be prepared from one or more sterile powders and/or granules havinga compound or salt of this invention and one or more of the carriers ordiluents mentioned for use in the formulations for oral administration.The powder or granule typically is added to an appropriate volume of asolvent (typically while agitating (e.g., stirring) the solvent) that iscapable of dissolving the powder or granule. Particular solvents usefulin the invention include, for example, water, polyethylene glycol,propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesameoil, benzyl alcohol, sodium chloride, and/or various buffers.

Emulsions for parenteral administration can be prepared by, for example,dissolving a compound or salt of this invention in any pharmaceuticallyacceptable solvent capable of dissolving the compound to form asolution; and adding an appropriate volume of a carrier to the solutionwhile stirring to form the emulsion. Solutions for parenteraladministration can be prepared by, for example, dissolving a compound orsalt of this invention in any pharmaceutically acceptable solventcapable of dissolving the compound to form a solution; and adding anappropriate volume of a carrier to the solution while stirring to formthe solution.

Suppositories for rectal administration can be prepared by, for example,mixing the drug with a suitable nonirritating excipient that is solid atordinary temperatures, but liquid at the rectal temperature and willtherefore melt in the rectum to release the drug. Suitable excipientsinclude, for example, cocoa butter; synthetic mono-, di-, ortriglycerides; fatty acids; and/or polyethylene glycols.

Every formulation or combination of components described or exemplifiedherein can be used to practice the invention, unless otherwise stated.

(i) Binding Agents

Binding agents include, but are not limited to, corn starch, potatostarch, or other starches, gelatin, natural and synthetic gums such asacacia, sodium alginate, alginic acid, other alginates, powderedtragacanth, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, carboxymethyl cellulose calcium, sodiumcarboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose,pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos.2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.Suitable forms of microcrystalline cellulose include, for example, thematerials sold as AVICEL-PH-101, AVICEL-PH-103 and AVICEL-PH-105(available from FMC Corporation, American Viscose Division, AvicelSales, Marcus Hook, Pa., USA). An exemplary suitable binder is a mixtureof microcrystalline cellulose and sodium carboxymethyl cellulose sold asAVICEL RC-581 by FMC Corporation.

(ii) Fillers

Fillers include, but are not limited to, talc, calcium carbonate (e.g.,granules or powder), lactose, microcrystalline cellulose, powderedcellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch,pre-gelatinized starch, and mixtures thereof.

(iii) Lubricants

Lubricants include, but are not limited to, calcium stearate, magnesiumstearate, mineral oil, electromagnetic radiation mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore,Md., USA), a coagulated aerosol of synthetic silica (marketed by DeaussaCo. of Plano, Tex., USA), CAB-β-SIL (a pyrogenic silicon dioxide productsold by Cabot Co. of Boston, Mass., USA), and mixtures thereof.

(iv) Disintegrants

Disintegrants include, but are not limited to, agar-agar, alginic acid,calcium carbonate, microcrystalline cellulose, croscarmellose sodium,crospovidone, polyacrilin potassium, sodium starch glycolate, potato ortapioca starch, other starches, pre-gelatinized starch, other starches,clays, other algins, other celluloses, gums, and mixtures thereof.

Tablets or capsules can optionally be coated by methods well known inthe art. If binders and/or fillers are used with a compound/bioconjugateof the invention, they are typically formulated as about 50 to about 99weight percent of the compound/bioconjugate. In one aspect, about 0.5 toabout 15 weight percent of disintegrant, and particularly about 1 toabout 5 weight percent of disintegrant, can be used in combination withthe compound. A lubricant can optionally be added, typically in anamount of less than about 1 weight percent of the compound/bioconjugate.Techniques and pharmaceutically acceptable additives for making solidoral dosage forms are described in Marshall, SOLID ORAL DOSAGE FORMS,Modern Pharmaceutics (Banker and Rhodes, Eds.), 7:359-427 (1979). Otherformulations are known in the art.

Liquid preparations for oral administration can take the form ofsolutions, syrups or suspensions. Alternatively, the liquid preparationscan be presented as a dry product for constitution with water or othersuitable vehicle before use. Such liquid preparations can be prepared byconventional means with pharmaceutically acceptable additives such assuspending agents (e.g., sorbitol syrup, cellulose derivatives orhydrogenated edible fats); emulsifying agents (e.g., lecithin oracacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethylalcohol or fractionated vegetable oils); and/or preservatives (e.g.,methyl or propyl-p-hydroxybenzoates or sorbic acid). The preparationscan also contain buffer salts, flavoring, coloring, perfuming andsweetening agents as appropriate. Preparations for oral administrationcan also be formulated to achieve controlled release of thecompound/bioconjugate. Oral formulations preferably contain 10% to 95%compound/bioconjugate. In addition, a compound/bioconjugate of thepresent invention can be formulated for buccal administration in theform of tablets or lozenges formulated in a conventional manner. Othermethods of oral delivery of compounds/bioconjugates of the inventionwill be known to the skilled artisan and are within the scope of theinvention.

Formulation 1

Hard gelatin capsules are prepared using the following ingredients:

TABLE F1 Ingredients (mg/capsule) Active Ingredient 250.0 Starch 305.0Magnesium stearate 5.0

The above ingredients are mixed and filled into hard gelatin capsules in560 mg quantities.

Formulation 2

A tablet formula is prepared using the following ingredients:

TABLE F2 Ingredients (mg/tablet) Active Ingredient 250.0 Cellulose,microcrystalline 400.0 Colloidal silicon dioxide 10.0 Stearic acid 5.0

The components are blended and compressed to form tablets, each weighing665 mg.

Formulation 3

A dry powder inhaler formulation is prepared containing the followingcomponents:

TABLE F3 Ingredients Weight % Active ingredient 5 Lactose 95

The active ingredient is mixed with the lactose and the mixture is addedto a dry powder inhaling appliance.

Formulation 4

Tablets, each containing 60 mg of active ingredient, are prepared asfollows:

TABLE F4 Ingredients milligrams Active ingredient 60.0 Starch 45.0Microcrystalline cellulose 35.0 Polyvinylpyrrolidone (as 10% solution inwater) 4.0 Sodium carboxymethyl starch 4.5 Magnesium stearate 0.5 Talc1.0 Total 150.0

The active ingredient, starch and cellulose are passed through a No. 20mesh U.S. sieve and mixed thoroughly. The solution ofpolyvinylpyrrolidone is mixed with the resultant powders which are thenpassed through a 16 mesh U.S. sieve. The granules as produced are driedat 50-60° C. and passed through a 16 mesh U.S. sieve. The sodiumcarboxymethyl starch, magnesium stearate, and talc, previously passedthrough a No. 30 mesh U.S. sieve, are then added to the granules which,after mixing, are compressed on a tablet machine to yield tablets eachweighing 150 mg.

Formulation 5

Capsules, each containing 80 mg of active ingredient are made asfollows:

TABLE F5 Ingredients milligrams Active ingredient 80.0 Starch 109.0Magnesium stearate 1.0 Total 190.0

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 190 mg quantities.

Formulation 6

Suppositories, each containing 225 mg of active ingredient, are made asfollows:

TABLE F6 Ingredients milligrams Active Ingredient 225 Saturated fattyacid glycerides to 2000

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2.0 g capacity and allowed to cool.

Formulation 7

Suspensions, each containing 50 mg of active ingredient per 5.0 ml doseare made as follows:

TABLE F7 Ingredients milligrams Active ingredient 50.0 mg Xanthan gum4.0 mg Sodium carboxymethyl cellulose (11%) Microcrystalline cellulose(89%) 50.0 mg Sucrose 1.75 g Sodium benzoate 10.0 mg Flavor q.v. Colorq.v. Purified water to 5.0 ml

The active ingredient, sucrose and xantham gum are blended, passedthrough a No. 10 mesh U.S. sieve, and mixed with a previously madesolution of the microcrystalline cellulose and sodium carboxymethylcellulose in water. The sodium benzoate, flavor, and color are dilutedwith some of the water and added with stirring. Sufficient water is thenadded to produce the required volume.

Formulation 8

Capsules, each containing 150 mg of active ingredient, are made asfollows:

TABLE F8 Ingredients milligrams Active ingredient 150.0 Starch 407.0Magnesium stearate 3.0 Total 560.0

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 20 mesh U.S. sieve, and filled into hardgelatin capsules in 560 mg quantities.

5f: Kits

Various embodiments of the present invention include kits. Such kits caninclude a compound/bioconjugate of the present invention, optionally oneor more ingredients for preparing a pharmaceutically acceptableformulation of the compound/bioconjugate, and instructions for use(e.g., administration). When supplied as a kit, different components ofa compound/bioconjugate formulation can be packaged in separatecontainers and admixed immediately before use. Such packaging of thecomponents separately can, if desired, be presented in a pack ordispenser device which can contain one or more unit dosage formscontaining the compound/bioconjugate. The pack can, for example,comprise metal or plastic foil such as a blister pack. Such packaging ofthe components separately can also, in certain instances, permitlong-term storage without losing activity of the components. Inaddition, if more than one route of administration is intended or morethan one schedule for administration is intended, the differentcomponents can be packaged separately and not mixed prior to use. Invarious embodiments, the different components can be packaged in onecombination for administration together.

It is further contemplated that the anti-angiogenesis agent compoundsand salts of this invention can be used in the form of a kit that issuitable for use in performing the methods described herein, packaged ina container. The kit can contain the anti-angiogenesis agent compound orcompounds and, optionally, appropriate diluents, devices or devicecomponents suitable for administration and instructions for use inaccordance with the methods of the invention. The devices can includeparenteral injection devices, such as syringes or transdermal patch orthe like. Device components can include cartridges for use in injectiondevices and the like. In one aspect, the kit includes a first dosageform including a anti-angiogenesis agent compound or salt of thisinvention and a second dosage form including another active ingredientin quantities sufficient to carry out the methods of the invention. Thefirst dosage form and the second dosage form together can include atherapeutically effective amount of the compounds for treating thetargeted condition(s).

In certain embodiments, kits can be supplied with instructionalmaterials. Instructions can be printed on paper or other substrate,and/or can be supplied as an electronic-readable medium, such as afloppy disc, mini-CD-ROM, CD-ROM, DVD-ROM, Zip disc, videotape, audiotape, and the like. Detailed instructions cannot be physicallyassociated with the kit; instead, a user can be directed to an Internetweb site specified by the manufacturer or distributor of the kit, orsupplied as electronic mail.

If desired, the emulsions or solutions described above for oral orparenteral administration can be packaged in IV bags, vials, or otherconventional containers in concentrated form, and then diluted with apharmaceutically acceptable liquid (e.g., saline) to form an acceptableanti-angiogenesis agent compound concentration before use.

Kits can include reagents in separate containers such as, for example,sterile water or saline to be added to a lyophilized active componentpackaged separately. For example, sealed glass ampules can containlyophilized superoxide dismutase mimetics and in a separate ampule,sterile water, sterile saline or sterile each of which has been packagedunder a neutral non-reacting gas, such as nitrogen. Ampules can consistof any suitable material, such as glass, organic polymers, such aspolycarbonate, polystyrene, ceramic, metal or any other materialtypically employed to hold reagents. Other examples of suitablecontainers include bottles that can be fabricated from similarsubstances as ampules, and envelopes that can consist of foil-linedinteriors, such as aluminum or an alloy. Other containers include testtubes, vials, flasks, bottles, syringes, and the like. Containers canhave a sterile access port, such as a bottle having a stopper that canbe pierced by a hypodermic injection needle. Other containers can havetwo compartments that are separated by a readily removable membrane thatupon removal permits the components to mix. Removable membranes can beglass, plastic, rubber, and the like.

Example 6 Synthesis of Quaternary Amine Compounds

Another aspect of the invention provides processes for the synthesis ofquaternary salt derivatives of the compounds described herein. Theprocesses comprise contacting a tertiary N-substituted compound withR²-(Leaving Group), wherein R² is as described in the context of (FX1)and (FX2), and X is a leaving group, to form a quaternary N-substitutedsalt.

(a) Synthesis of Compounds Comprising Formula (FX11) and (FX12)

In one embodiment, a quaternary salt compound comprising Formula (FX11)or (FX12) is synthesized from a compound comprising Formula (FX9) or(FX10). For purposes of illustration, Reaction Scheme 6-1 depictsproduction of a compound comprising Formula (FX11) in accordance withone aspect of the invention:

For purposes of illustration, Reaction Scheme 6-2 depicts production ofa compound comprising Formula (FX12) in accordance with one aspect ofthe invention:

(b) Reaction Mixture

Synthesis commences with formation of a reaction mixture by combining acompound comprising Formulas (FX1) or (FX2) with R²-(Leaving Group).Thus, for example, R²-(Leaving Group) may be a methyl, ethyl, propyl,allyl, cyclopropyl, cyclopropylmethyl, propargyl, or benzyl halide oranother leaving group. Non-limiting examples of R²-(Leaving Group)include methyl bromide, methyl chloride, allyl iodide, cyclopropylmethylbromide, dimethyl sulfate, diethyl sulfate, di(cyclopropylmethyl)sulfate, methyl fluorosulfonate, trimethyloxonium fluoroborate,triethyloxonium fluoroborate, trimethyloxonium hexachloroantimonate, npropyl or n-octyl trifluoromethane sulfonate, trimethyloxoniumhexafluorophosphate, methyl trifluoromethane sulfonate, methyl iodide,and allyl trifluoromethanesulfonate. In a preferred embodiment,R²-(Leaving Group) is methyl bromide.

The mole-to-mole ratio of the compound comprising Formulas (FX1) or(FX2) to R²-(Leaving Group) can vary. In general, the mole-to-mole ratioof the compound comprising Formulas (FX1) or (FX2) to R²-(Leaving Group)may range from about 1:1 to about 1:2. In some embodiments, themole-to-mole ratio of the compound comprising Formulas (FX1) or (FX2) toR²-(Leaving Group) may be about 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5,1:1.6, 1:1.7, 1:1.8, 1:1.9, or 1:2. In preferred embodiments themole-to-mole ratio of the compound comprising Formulas (FX1) or (FX2) toR²-(Leaving Group) may range from about 1:1 to about 1:1.5. Furthermore,the rate of addition of R²-(Leaving Group) to the compound comprisingFormulas (FX1) or (FX2) may vary. Typically, the rate of addition ofR²-(Leaving Group) may range from 0.002 to about 0.02 equivalents ofR²-(Leaving Group) per minute per equivalent of the compound comprisingFormulas (FX1) or (FX2) in the reaction mixture.

The reaction mixture, as detailed herein, also comprises a solventsystem. In general, the solvent system is anhydrous. That is, thesolvent system comprises less than about 0.5% of water by weight,typically less than about 0.2% of water by weight, and in someembodiments, less than about 0.05% of water by weight. The solventsystem typically comprises an aprotic solvent. Non-limiting examples ofsuitable aprotic solvents include acetonitrile, 1,4-dioxane,N,N-dimethylacetamide (DMAC), N,N-dimethylformamide (DMF),N-methyl-2-pyrrolidinone (NMP), hexamethylphosphoramide, (HMPA), andcombinations thereof. In a preferred embodiment, the aprotic solvent isN-methyl-2-pyrrolidinone (NMP). The solvent system may additionallycomprise an organic solvent such as ether, hydrocarbon, toluene,benzene, halobenzene, xylenes, or combinations thereof. In general, thesolvent system comprises at least about 50% of the aprotic solvent byweight, at least about 75% of the aprotic solvent by weight, or at leastabout 90% of the aprotic solvent by weight.

The amount of solvent system in the reaction mixture may vary.Typically, the weight-to-weight ratio of solvent system to the compoundcomprising Formulas (FX1) or (FX2) may range from about 1.5:1 to about2.0:1. In some embodiments, the weight-to-weight ratio of solvent systemto the compound comprising Formulas (FX1) or (FX2) may be about 1.5:1.1.55:1, 1.6:1, 1.65:1, 1.7:1, 1.75:1, 1.8:1, 1.85:1, 1.9:1, 1.95:1, or2.0:1.

In some embodiments, X is a hydroxy group in the compound comprisingFormula (FX1) or (FX2), wherein reaction with R²-(Leaving Group) mayyield undesirable alkyloxymorphinan compounds. To prevent such sidereactions, an acid may be added to the reaction mixture to suppressionization of the hydroxy group. Suitable acids include strong mineralor organic acids. For example, the acid may be a carboxylic acid, aphosphonic acid, a sulfonic acid, or a mixture thereof. Alternatively, asmall amount of a preformed alkaloid acid salt may be added to itsalkaloid base in order to suppress ionization of the alkaloid base; forexample, naltrexone hydrobromide may be added to naltrexone base. By wayof further example, the acid may be HBr, HCl, H₂SO₄, NaHSO₄, NaH₂PO₄, orNa₂HPO₄. In preferred embodiments, the acid may be HBr gas or HCl gas.Preferably, the acid is also anhydrous. That is, the acid may compriseless that about 0.5% of water by weight, less than about 0.2% of waterby weight, or more preferably, less than about 0.05% of water by weight.Those of skill in the art will also appreciate that the 3-hydroxy of thecompound comprising Formulas (FX1) or (FX2) may be protected with ahydroxy protecting group prior to the reaction of the invention.

(c) Reaction Conditions

The quaternization reaction may be carried out over a wide range oftemperatures and pressures. Typically, the reaction will be carried outat a temperature that ranges from about room temperature (i.e., about25° C.) to about 90° C. In preferred embodiments, the temperature of thereaction may range from about 55° C. to about 85° C. In someembodiments, the temperature of the reaction may be about 55°, 60°, 65°,70°, 75°, 80°, or about 85° C. In general, the reaction will beconducted at a pressure of no more than about 4 atmospheres. Inpreferred embodiments, the pressure of the reaction may range from about1 to about 2 atmospheres. In other embodiment, the reaction may occur atatmospheric pressure.

The duration of the reaction can and will vary. For example, thereaction may be allowed to proceed from about several hours to aboutseveral days. Typically, however, the reaction is allowed to proceed fora sufficient period of time until the reaction is complete, asdetermined by means well known to those of skill in the art. In thiscontext, a “completed reaction” generally means that the final reactionmixture contains a significantly diminished amount of the reactants anda significantly increased amount of the products of Reaction Schemes 1and 2 compared to the amounts of each present at the beginning of thereaction. Typically, the amount of the reactant remaining in the finalreaction mixture may be less than about 5%, and preferably less thanabout 1%.

When the reaction is completed, the reaction mixture is generally cooledto at least about room temperature, such that the reaction product maybe isolated. In some embodiments, a solvent of lower polarity in whichthe reaction product is not soluble may be added to the cooled reactionmixture to facilitate precipitation of the quaternary reaction productwhile leaving the unreacted tertiary substrate in solution. Examples ofsuitable solvents include, but are not limited to acetone, chloroform,dichloromethane, ethyl acetate, propyl acetate, methyl ethyl ketone,methyl butyl ketone, ether, t-butylmethylether, 2-methyltetrahydrofuran,hydrocarbon, toluene, benzene, chlorobenzene, bromobenzene, and mixturesthereof. The reaction mixture may be optionally cooled further to about0° C. to about 5° C. The precipitated product generally is separatedfrom the remaining reaction mixture by filtration, and is washed anddried to produce the final product. The yield of the final producttypically will range form about 50% to about 99%. In some embodiments,the yield of the final product may be at least about 50, 55, 60, 65, 70,75, 80, 85, 90, or 95%.

In embodiments in which the Leaving Group is a halide, the final productcomprises a halide anion. This anion may be exchanged by treating thecompound with a protic acid, thereby replacing the halide ion withanother anion such as nitrate, sulfate, phosphate, or another halideion.

(d) Synthesis of R-3-Acetoxy-(+)-Naltrexone Methobromide

The following reaction scheme depicts the synthesis of3-acetoxy-M-naltrexone methobromide:

A solution of 3-acetoxy-(+)-naltrexone in 1-methyl-3-pyrrolidinone (NMP)(732.2 g of 30% wt/wt solution, 0.57 moles) may be added to a 1-L,5-neck, jacketed pressure reactor equipped with a polished glassstirring shaft, mechanical stirrer, reflux condenser, pressure manifold,thermowell, and ⅛″ (id) addition line. Methyl bromide (MeBr, 107.9 g,1.14 moles) then may be added subsurface through the addition line withvigorous stirring of the solution over a 1-hour period. The amount ofMeBr is determined by a difference in the initial and final weights of aMeBr lecture bottle. During the addition, the temperature of thereaction mixture may increase to about 33° C. (yellow solution) with amaximum pressure of 3-4 psi. After the appropriate amount of MeBr isadded, the reactor headspace may be evacuated and re-pressurized withMeBr (to about 2 psi) twice before heating to 60° C. At 60° C., apressure of 2-4 psi may be observed. The reaction mixture may be stirredovernight (15 hours) with no pressure remaining over the yellowsolution. Aqueous hydrogen bromide (HBr, 1.0 equiv, 0.57 moles, 96.58 gof 48 wt. %) may be added slowly at 60° C. over a 30-minute period. Thereactor may be vented into NMP in order to trap gaseous methyl bromidethat is generated during the HBr addition. During the addition, thereaction temperature may increase to about 64° C. The reactiontemperature then may be increased to 80° C. over a 1.5-hour period,wherein evolution of methyl bromide ceases. The mixture may be stirredat 80° C. for 2 hours, wherein precipitation occurs. After 5 hours at80° C., the slurry may be analyzed by HPLC. Preferably, only a minoramount of 3-acetoxy-(+)-naltrexone methobromide remains in the slurry(<0.5% by area). The mixture then may be transferred to a 2-L three-neckround-bottomed flask equipped with a glass stirring shaft, mechanicalstirrer, reflux condenser, and thermocouple under a nitrogen atmosphere.The mixture may be cooled to about 56° C. and methanol (512.5 g, 1.0 wtequiv. based on the amount of NMP charged) may be added quickly tofacilitate crystallization of methobromide salts. The slurry then may becooled to about 20° C. over a 30-minute period and then to about 5-10°C. in an ice bath. The slurry may be stirred for 1 hour at 5-10° C.,filtered, and the product washed with cold methanol (319 mL, 1.45 mL/gof starting material to afford product as a white solid (e.g., about236.1 g of product; 87.2%). The crude product may be analyzed by HPLC(e.g., it may contain 88.54% R and 1.47% S diastereomers).

(e) Synthesis of R-(+)-Naltrexone Methobromide

The following reaction scheme depicts the synthesis of (+)-naltrexonemethobromide:

Fresh anhydrous 1-methyl-2-pyrrolidinone (50 mL) may be added to a3-necked 250 mL flask fitted with a thermocouple, addition funnel,condenser, and a mechanical stirrer under a sweep of dry nitrogen. Thesolution may be heated to 55° C. The addition funnel may be replacedwith a powder funnel and anhydrous (+)naltrexone base (39.5 grams) addedwith stirring. After the funnel is “washed down” with 10 mL ofadditional 1-methyl-2-pyrrolidinone, the temperature may be is adjustedto 55-58° C. and the addition funnel replaced on the flask. Separately,10 mL of anhydrous 1-methyl-2-pyrrolidinone may be cooled in a graduatedcylinder, and methyl bromide gas may be condensed in a lecture bottleusing an ice bath and 10 mL measured out as a liquid into another coldgraduated cylinder. The cold methyl bromide liquid and1-methyl-2-pyrrolidinone may be combined and mixed. The methyl bromidesolution may be poured into the addition funnel and added dropwise tothe (+) naltrexone solution under a slow sweep of dry nitrogen. Thetemperature of the solution may increase to about 66° C. The reactiontemperature and time may be set at 62.5° C. for nine hours. After anhour, a fine white suspension of R-(+)-naltrexone methobromide may beginto form. At the end of nine hours the heating may be discontinued andthe mixture may be allowed to cool to room temperature with stirringovernight. Acetone (75 mL) may be poured into the suspension tofacilitate the precipitation of product. The slurry may be cooled to icebath temperature and stirred. The product may be recovered by vacuumfiltration and washed with 25 mL of additional acetone. The product maybe dried to a constant weight in a vacuum oven set at 60° C. The yieldof the unpurified salts may be about 31.8 g.

(f) Synthesis of R-(+)-Naloxone Methobromide

Anhydrous 1-methyl-2-pyrrolidinone (5 mL) may be added to a 25 mL flaskfitted with a condenser and stirring bar under a sweep of dry nitrogen.Anhydrous (+)naloxone base (4.11 grams) may be added with stirring.Methyl bromide gas may be condensed in a lecture bottle using an icebath and 0.5 mL measured out as a liquid in another cold graduatedcylinder. The methyl bromide may be poured into the naltrexone basesuspension under a slow sweep of dry nitrogen. The reaction temperatureand time may be set at 60° C. for ten hours. At the end of ten hours theheating may be discontinued and the mixture allowed to cool to roomtemperature with stirring overnight (or longer). Acetone (10 mL) may beadded to the suspension to facilitate the precipitation of the product.The slurry may be cooled to ice bath temperature with stirring. Theproduct may be recovered by vacuum filtration and washed with additionalacetone. The product may be dried in a vacuum oven set at 60° C. for twohours. For example, 2.89 grams of the crude product may be recovered.Recrystallization from methanol/water (20 30 mL, 8:2) may yield 2.43grams of a white crystalline salt.

(g) Synthesis of R-(+)-Nalfurafine Methoiodide

(+)-Nalfurafine (2.0 g, 4.3 mmol), ethyl acetate (60 mL), methanol (6mL), and methyl iodide (1.3 mL) may be placed together in a sealedreactor. The reactor contents may be stirred at 100° C. for about fourdays. Methanol (60 mL) may be added to the reaction solution tofacilitate precipitation of the product. The precipitated solid may bedissolved and concentrated. Distilled water (400 mL) may be added to theresulting residue. This aqueous solution may be washed with chloroform(7×100 mL). The water phase may be concentrated. The resulting residuemay be recrystallized from ethyl acetate-methanol. The resulting crystalmay be dissolved in distilled water (500 mL). This aqueous solution maybe washed with chloroform (3×100 mL). The water phase may beconcentrated. The resulting residue may be recrystallized three timesfrom methanol. As an example, 102 mg of the product, R-(+)-nalfurafinemethoiodide, may be obtained.

(h) Synthesis of R-(+)-Oxycodone Cyclopropylmethobromide

The following reaction scheme depicts the synthesis of (+)-oxycodonecyclopropylmethobromide:

A reactor was charged with (+)-oxycodone (0.5 g, 1.595 mmol) and 2 mL of1-methyl-3-pyrrollidinone (NMP). The reactor was flushed with nitrogenand the mixture was kept under nitrogen throughout the reaction.Cyclopropylmethylbromide (0.2 mL, 1.3 eq) was added and the reactionmixture was heated to 68° C. for 3 h. Samples were removed at 1 h, 2 h,and 3 h for analysis; the reaction was complete after 2 h. The reactionmixture was cooled to room temperature. Acetone (4 mL) was added and themixture was stirred at 20° C. for 1 h. The solids were filtered andwashed with acetone (3×2 mL); 0.2 g of solid was recovered. The filtrateand acetone wash were pumped down (and washed with water) to giveanother 0.22 g of solid. The solids were combined, charged with water (5mL, stirred at rt for 2 h), and filtered. The solid was washed withwater (3×2 mL), dried in a vacuum oven at 65° C. for 18 h to give 0.36 gof white solid.

Example 7 Synthesis of N-Oxide Compounds

Another aspect of the invention provides processes for the synthesis ofN-oxide derivatives of the compounds described herein. The processescomprise contacting a tertiary N-substituted compound with an oxidizingagent, to form a quaternary N-oxide.

(a) Synthesis of Compounds Comprising Formula (FX3) and (FX4)

In one embodiment, a quaternary salt compound comprising Formula (FX3)or (FX4) is synthesized from a compound comprising Formula (FX9) or(FX10). For purposes of illustration, Reaction Scheme 7-1 depictsproduction of a compound comprising Formula (FX3) in accordance with oneaspect of the invention:

For purposes of illustration, Reaction Scheme 7-2 depicts production ofa compound comprising Formula (FX4) in accordance with one aspect of theinvention:

(b) Reaction Mixture

The process of the invention commences with formation of a reactionmixture by combining a compound comprising Formulas (FX1) or (FX2) withan oxidizing agent A variety of oxidizing agents are suitable for use inthe process of the invention. Examples of oxidizing agents that may beused include, but are not limited to tungsten(VI) oxide, chromium oxide,dichromate, copper oxide, nickel oxide, cobalt oxide, silver oxide,oxides of mercury, oxides of lead, selenium oxide, ruthenium oxide,hydrogen peroxide, peroxysulfate, peroxyacetic acid,3-chloroperoxybenzoic acid, RCO₃H, wherein R is selected from the groupconsisting of hydrogen, an alkyl, a substituted alkyl, an aryl, asubstituted aryl, dichromates (e.g., ammonium dichromate, potassiumdichromate, sodium dichromate, and the like); bromates (e.g., bariumbromate, magnesium bromate, potassium bromate, sodium bromate, and thelike); chlorates (e.g., ammonium chlorate, barium chlorate, calciumchlorate, potassium chlorate, sodium chlorate, and the like); chlorites(e.g., copper chlorite, lead chlorite, potassium chlorite, sodiumchlorite, and the like); chloroisocyanuric acids (e.g.,trichloroisocyanuric acid, and the like); chromates (e.g., potassiumchromate, and the like); chromium oxides (e.g., chromic anhydride(chromium trioxide)); dichromates (e.g., sodium dichromate, potassiumdichromate, and the like); hydrogen peroxide; hypobromites (e.g., sodiumhypobromite, and the like); hypochlorites (e.g., calcium hypochlorite,potassium hypochlorite, sodium hypochlorite, and the like); hypoiodites(e.g., sodium hypoiodite, potassium hypoiodite, and the like); inorganicperoxides (e.g., barium peroxide, calcium peroxide, cesium peroxide,lithium peroxide, magnesium peroxide, potassium peroxide, rubidiumperoxide, sodium peroxide, strontium peroxide, and the like); iodates(e.g., calcium iodate, potassium iodate, sodium iodate, zinc iodate, andthe like); iodine oxides (e.g., diiodine pentaoxide, and the like); leadoxides (e.g., lead dioxde, and the like); manganese dioxide; nitrates(e.g., ammonium nitrate, ammonium cerium nitrate, barium nitrate,potassium nitrate, silver nitrate, sodium nitrate, and the like); nitricacid; nitrites (e.g., potassium nitrite, sodium nitrite, and the like);perchlorates (e.g., ammonium perchlorate, potassium perchlorate, sodiumperchlorate, and the like); periodates (e.g., potassium periodate,sodium periodate, and the like); periodic acids (e.g., metaperiodicacid, and the like); permanganates (e.g., ammonium permanganate,magnesium permanganate, potassium permanganate, sodium permanganate, andthe like); peroxoborates (e.g., ammonium peroxoborate, and the like);perchloric acid; peroxodisulfates (e.g., ammonium peroxodisulfates,potassium peroxydisulfate, and the like); peroxyacids (e.g.,peroxyacetic acid, peroxybenzoic acid, peroxyformic acid,trifluoroperacetic acid, and the like); organic peroxides (e.g., benzoylperoxide, and the like); tetroxides (e.g., osmium tetroxide, rutheniumtetroxide, and the like); dimethyldioxirane; and oxygen. As the oxygensource, air may also be used. In an exemplary embodiment the oxidizingagent is hydrogen peroxide.

The mole-to-mole ratio of the compound comprising Formulas (FX1) or(FX2) to oxidizing agent can vary. In general, the mole-to-mole ratio ofthe compound comprising Formulas (FX1) or (FX2) to oxidizing agent mayrange from about 1:1 to about 1:20. In some embodiments, themole-to-mole ratio of the compound comprising Formulas (FX1) or (FX2) tooxidizing agent may be about 1:1, 1:1.1, 1:1.5, 1:2, 1:3, 1:4, 1:5, 1:6,1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18,1:19, or 1:20. In preferred embodiments the mole-to-mole ratio of thecompound comprising Formulas (FX1) or (FX2) to oxidizing agent may rangefrom about 1:10 to about 1:15.

The reaction mixture, as detailed herein, generally also comprises asolvent. Those of skill in the art will appreciate that they solventutilized will depend upon a variety of factors, including the chemicalnature of the starting compound. In some embodiments, the solvent may bea protic solvent. Non-limiting suitable examples of protic solventsinclude, but are not limited to, methanol, ethanol, isopropanol,n-propanol, isobutanol, n-butanol, s-butanol, f-butanol, formic acid,acetic acid, water, and combinations thereof. In an exemplaryembodiment, the protic solvent is methanol. In other embodiments, thesolvent may be an aprotic solvent. Non-limiting examples of suitableaprotic solvents include acetone, acetonitrile, N,N-dimethylformamide(DMF), dimethyl sulfoxide (DMSO), N,N-dimethylpropionamide,1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU),1,3-dimethyl-2-imidazolidinone (DMI), N,N-dimethylacetamide (DMAC),N-methyl-2-pyrrolidinone (NMP), ethyl acetate, ethyl formate, ethylmethyl ketone, formamide, hexachloroacetone, hexamethylphosphoramide,methyl acetate, N-methylacetamide, N-methylformamide, methylenechloride, nitrobenzene, nitromethane, propionitrile, sulfolane,tetramethylurea, toluene, trichloromethane, xylenes, and combinationsthereof. In still other embodiments, the solvent may be an organicsolvent. Suitable organic solvents include, but are not limited to,alkane and substituted alkane solvents (including cycloalkanes),aromatic hydrocarbons, esters, ethers, combinations thereof, and thelike. Specific organic solvents that may be employed, include, forexample, acetonitrile, benzene, butyl acetate, t-butyl methylether,chlorobenzene, chloroform, chloromethane, cyclohexane, dichloromethane,dichloroethane, ethyl acetate, diethylene glycol, fluorobenzene,heptane, hexane, isopropyl acetate, pentyl acetate, n-propyl acetate,tetrahydrofuran, toluene, and combinations thereof. In additionalembodiments, the solvent may comprise a mixture of protic, aprotic,and/or organic solvents as delineated above.

The amount of solvent in the reaction mixture may vary. Typically, theweight-to-weight ratio of solvent to the compound comprising Formulas(FX1) or (FX2) may range from about 2:1 to about 100:1, preferably fromabout 3:1 to about 30:1, or more preferably from about 5:1 to about15:1. In some embodiments, the weight-to-weight ratio of solvent to thecompound comprising Formulas (FX1) or (FX2) may be about 1:5, 1:6, 1:7,1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, or 1:15.

(c) Reaction Conditions

In general, the oxidation reaction may be conducted at a temperaturethat ranges from about −10° C. to about 120° C. for a period of timethat is sufficient to convert a substantial portion of the reactants toproducts. In one embodiment, the temperature of the reaction may rangefrom about −10° C. to about 80° C. In another embodiment, thetemperature may range from about −10° C. to about 50° C. In stillanother embodiment, the temperature of the reaction may range from about−5° C. to about 30° C. In a further embodiment, the temperature of thereaction may be about room temperature (˜25° C.). The reaction ispreferably performed under ambient pressures and preferably in an inertatmosphere (e.g., nitrogen or argon).

Typically, the reaction is allowed to proceed for a sufficient period oftime until the reaction is complete, as determined by chromatography(e.g., TLC or HPLC). In this context, a “completed reaction” generallymeans that the reaction mixture contains a significantly diminishedamount of reactants and a significantly increased amount of productscompared to the amounts of each present at the beginning of thereaction. Typically, the amount of reactants remaining in the reactionmixture may be less than about 5%, and preferably less than about 1%.

Upon completion of the reaction, the reaction mixture may be cooled toabout 4° C. or less. Once the solution has cooled, filtration may beperformed (for example by use of a celite plug) to remove impurities.The product may be isolated by phase extraction, liquid chromatography,crystallization, or other means familiar to those of skill in the art.The final product may washed and dried, and analyzed by HPLC, UPLC, MS,NMR, IR, or TGA. The yield of the compound comprising Formula (II) orFormula (IV) may vary. Typically, the yield of the compound may rangefrom about 60% to about 99%, and more specifically from about 70% toabout 80%.

(d) Synthesis of (+)-Hydrocodone N-Oxide

The following scheme depicts the synthesis of (+)-hydrocodone N-oxidefrom (+)-hydrocodone.

Hydrogen peroxide (2.5 mL, 50% w/v, 36.8 mmol, 11 eq.) was addeddrop-wise to a cooled solution of (+)-hydrocodone (1.0 g, 3.34 mmol, 1.0eq.) in 10 mL of methanol (MeOH) in an ice bath. The reaction mixturewas gradually warmed to room temperature overnight. High phase liquidchromatography (HPLC) analysis indicated the reaction was done.Approximately 50 mg MnO₂ was added to the reaction to decompose theexcess hydrogen peroxide. Bubbling was observed after adding MnO₂. Afterthe bubbling stopped, the reaction was filtered through celite and thesolid residue was washed with methanol (3 mL×3). After removal of thevolatiles from the combined filtrates on a rotoevaporator unit, a greysolid remained. The grey solid was added to 30 mL brine, the resultingsuspension was cooled to approximately 0° C. in an ice bath, and the pHwas adjusted to 1 with 6 N HCl. The product was extracted withdichloromethane (3×20 mL) and the combined organic phases were driedover anhydrous magnesium sulfate. After removing volatiles of the driedorganic phase by means of the rotoevaporator, a white solid was left,0.78 g, a yield of 74%, with purity=98.7%. Two peaks were observed onultra pressure liquid chromatography (UPLC) analysis, the smaller peakhad 5.17% ratio and the bigger peak had 94.83% ratio. Both peaks hadM+1=316.3 and fragmentation pattern on LC-MS analysis.

(e) Synthesis of (+)-Hydrocodone N-Oxide and Isolation of Isomers

The following reaction scheme depicts the oxidation of (+)-hydrocodoneto form (+)-hydrocodone N-oxide.

(+)-Hydrocodone N-oxide may be prepared in high yield by N-oxidation of(+)-hydrocodone base with hydrogen peroxide in the presence of a proticsolvent. Accordingly, a suspension of 4.348 g (+)-hydrocodone base(14.52 mmol) in 50 mL of methanol may be slowly mixed with 19 mL of 30%hydrogen peroxide (H₂O₂) (186 mmol, 12.8 eq.) at room temperature. Thesuspension may be stirred overnight at room temperature under nitrogen.The completeness of the reaction may be monitored by TLC analysis (e.g.,CHCl₃:MeOH:NH₄OH, 80:20:1). The starting material should be consumed andtwo products may be detected (major product, R_(f) 0.44, and minorproduct, R_(f) 0.25). The reaction mixture may be was cooled in anice-water bath and the quenched with manganese dioxide (MnO₂). Afterfiltration though a plug of celite followed by concentration, the crudereaction mixture may be subjected to flash column chromatography(CHCl₃:MeOH:NH₄OH gradient, from 100:0:1 to 95:5:1) such that bothproducts are isolated. By mass spectroscopy (MS) analysis, both major(3.7 g) and minor (230 mg) products have the same molecular weight andsimilar fragmentation patterns consistent for the N-oxide isomers. HPLCmay be use to determine that both peaks (major at 6.65 min, minor at4.37 min) are chromatographically pure. Nuclear magnetic resonance (NMR)may be used to show that the minor product could be a mixture of two orthree compounds. NMR analysis may show that the major product is the (+)hydrocodone N-oxide with an axial N—O bond. The major isomer may bepurified by flash column chromatography (CHCl₃:MeOH:NH₄OH gradient, from100:0:1 to 95:5:1) with four fractions being collected (F1, 97.55% area;F2, 98.05% area; F3, 97.51% area and F4, 98.19% area). Fractions 2 and 4may be combined, and after removal of the solvent, 1.45 g of the majorproduct may be obtained (C1, 98.36% area) as an off-white foam.Fractions 1 and 3 may be combined, and after solvent removal, 1.26 g ofproduct may be obtained (C2) as an off-white foam. C1 may be furtheranalyzed and verified by infrared spectroscopy (IR), thermogravimetricanalysis (TGA), NMR, HPLC, and MS methods.

STATEMENTS REGARDING INCORPORATION BY REFERENCE AND VARIATIONS

All references throughout this application, for example patent documentsincluding issued or granted patents or equivalents; patent applicationpublications; and non-patent literature documents or other sourcematerial; are hereby incorporated by reference herein in theirentireties, as though individually incorporated by reference, to theextent each reference is at least partially not inconsistent with thedisclosure in this application (for example, a reference that ispartially inconsistent is incorporated by reference except for thepartially inconsistent portion of the reference).

U.S. patent application Ser. No. 12/710,383, filed on Feb. 23, 2010,U.S. patent application Ser. No. 12/710,379, filed on Feb. 23, 2010, PCTInternational Application No. PCT/US10/24963, filed on Feb. 23, 2010,and PCT International Application No. PCT. US10/24961, filed on Feb. 23,2010, are each hereby incorporated by reference in its entirety andgenerally relate to methods of synthesizing and derivatizingmorphinanium n-oxides, morphinanium quaternary salts and stereoisomersthereof.

U.S. Provisional Patent Application No. 61/226,015, filed on Jul. 16,2009, and U.S. Provisional Patent Application No. 61/286,877, filed onDec. 16, 2009, are each hereby incorporated by reference in its entiretyand generally relate to (+) morphinan compounds and related methods ofsynthesizing and using (+) morphinan compounds, for example intherapeutic methods.

PCT International Application No. PCT/US2007/025263, filed on Dec. 10,2007 and published on Jun. 19, 2008 as International Publication No.WO/2008/073390; PCT International Application No. PCT/US2007/025262,filed on Dec. 10, 2007 and published on Jun. 19, 2008 as InternationalPublication No. WO/2008/073389; and PCT International Application No.PCT/US2005/038140, filed on Oct. 21, 2005 and published on May 18, 2006as International Publication No. WO/2006/052430 are each herebyincorporated by reference in its entirety and generally relate to thesynthesis and derivatization of isoquinolines, for example by totalopiate synthesis and chiral reduction techniques.

U.S. patent application Ser. No. 12/316,862, filed on Dec. 17, 2008 andpublished on Jun. 18, 2009 as U.S. Patent Publication No. 2009/0156,818;and U.S. patent application Ser. No. 12/316,846, filed on Dec. 17, 2008and published on Jun. 18, 2009 as U.S. Patent Publication No.2009/0156,816 are each hereby incorporated by reference in its entiretyand generally relate to the synthesis and derivatization of (+)morphinan compounds, for example using the natural product Sinomenine.

U.S. patent application Ser. No. 12/485,200, filed on Jun. 16, 2009 andpublished on Dec. 17, 2009 as U.S. Patent Publication No. 2009/0312,552;is hereby incorporated by reference in its entirety and generallyrelates to a chemical process to make a beta epimer for b-naltrexolpreparation.

U.S. patent application Ser. No. 12/316,861, filed on Dec. 17, 2008 andpublished on Jun. 18, 2009 as U.S. Patent Publication No. 2009/0156,817;is hereby incorporated by reference in its entirety and generallyrelates to process chemistry to make bases such as, the (+)buprenorphinebase, which may be further derivatized (e.g., oxidize or alkylate) usingprocesses disclosed herein and known in the art.

PCT International Application No. PCT/US2008/068103, filed on Jun. 25,2008 and published on Jan. 15, 2009 as International Publication No.WO/2009/009292; is hereby incorporated by reference in its entirety andgenerally relates to the preparation of naltrexone methobromide, forexample of morphic crystalline forms of naltrexone methobromide.

PCT International Application No. PCT/US2008/062413, filed on May 2,2008 and published on Nov. 13, 2008 as International Publication No.WO/2008/137672; is hereby incorporated by reference in its entirety andgenerally relates to preparation of a 6-alpha epimer hydroxymorphinanand synthesis of nalbuphine.

PCT International Application No. PCT/US07/019,489, filed on Sep. 6,2007 and published on Mar. 27, 2008 as International Publication No.WO/2008/036172; is hereby incorporated by reference in its entirety andgenerally relates to chemistry and synthetic processes that bridgeintermediates from benzyl isoquinolines to a primary morphinanstructure, including description of derivatization and transformation ofthe morphinan structure to product opiates or their mirror isomers.

PCT International Application No. PCT/US06/006285, filed on Feb. 23,2006 and published on Sep. 21, 2006 as International Publication No.WO/2006/098855 is hereby incorporated by reference in its entirety andgenerally relates to processes for making and derivatizing primarymorphinan compounds.

PCT International Application No. PCT/US03/035463, filed on Nov. 5, 2003and published on May 27, 2004 as International Publication No.WO/2004/043964; and PCT International Application No. PCT/US08/003,070,filed on Mar. 6, 2008 and published on Sep. 12, 2008 as InternationalPublication No. WO/2008/109156; are each hereby incorporated byreference in its entirety and generally processes and syntheticapproaches for preparing quaternary alkaloids salts.

PCT International Application No. PCT/US05/029437, filed on Aug. 17,2005 and published on Mar. 2, 2006 as International Publication No.WO/2006/023669 is hereby incorporated by reference in its entirety andgenerally relates to processes and synthetic approaches for making andderivatizing levorphanol.

U.S. Pat. No. 4,521,601 issued on Jun. 4, 1985 is hereby incorporated byreference in its entirety and generally relates to methods of making andderivatizing (+)morphinan alkaloid compounds.

Each of the following references are hereby incorporated by referenceits entirety and relates generally to differences in binding to theopiate receptor of the opiate isomers:

-   (1) Studies in the (+)-morphinan series. 5. Synthesis and biological    properties of (+)-naloxone. Iijima, Ikuo; Minamikawa, Junichi;    Jacobson, Arthur E.; Brossi, Arnold; Rice, Kenner C.; Klee,    Werner A. Lab. Chem., Natl. Inst. Arthritis, Metab. Dig. Dis.,    Bethesda, Md., USA. Journal of Medicinal Chemistry (1978), 21(4),    398-400;-   (2) “Unnatural alkaloids”. Brossi, Arnold. Natl. Inst. Arthritis    Metab. Dig. Dis., Natl. Inst. Health, Bethesda, Md., USA. Pure and    Applied Chemistry (1979), 51(4), 681-8;-   (3) Efficient syntheses and biological evaluation of novel    (+)-morphinans. Brossi, Arnold; Kerekes, Peter; Chi-shen, Chang.    Lab. Chem., Med. Chem. Sect., Natl. Inst. Arthritis, Diabetes    Digest. Kidney Dis., Bethesda, Md., USA. Studies in Organic    Chemistry (Amsterdam) (1985), 20(Nat. Prod. Chem.), 15-24; and-   (4) Stereospecific and nonstereospecific effects of (+)- and    (−)-morphine: Evidence for a new class of receptors? Jacquet, Yasuko    F.; Klee, Werner A.; Rice, Kenner C.; Iijima, Ikuo; Minamikawa,    Junichi. New York State Res. Inst. Neurochem, Drug Addict., Ward's    Island, N.Y., USA. Science (Washington, D.C., United States) (1977),    198(4319), 842-5.

PCT International Application No. PCT/US08/064412, filed on May 21, 2008and published on May 28, 2009 as International Publication No.WO/2009/067275; PCT International Application No. PCT/US07/85458, filedon Nov. 21, 2007 and published on May 29, 2008 as InternationalPublication No. WO/2008/064351; and U.S. patent application Ser. No.11/944,242, filed on Nov. 21, 2007 and published on Aug. 28, 2008 asU.S. Publication No. 20080207669 are each hereby incorporated byreference in its entirety and generally relate to morphinanium oxidecompounds, including (−) enantiomers or naturally derived morphinaniumoxide compounds.

The terms and expressions which have been employed herein are used asterms of description and not of limitation, and there is no intention inthe use of such terms and expressions of excluding any equivalents ofthe features shown and described or portions thereof, but it isrecognized that various modifications are possible within the scope ofthe invention claimed. Thus, it should be understood that although theinvention has been specifically disclosed by preferred embodiments,exemplary embodiments and optional features, modification and variationof the concepts herein disclosed can be resorted to by those skilled inthe art, and that such modifications and variations are considered to bewithin the scope of this invention as defined by the appended claims.The specific embodiments provided herein are examples of usefulembodiments of the invention and it will be apparent to one skilled inthe art that the invention can be carried out using a large number ofvariations of the devices, device components, methods steps set forth inthe present description. As will be apparent to one of skill in the art,methods and devices useful for the present methods can include a largenumber of optional composition and processing elements and steps.

When a group of substituents is disclosed herein, it is understood thatall individual members of that group and all subgroups, including anyisomers, enantiomers, and diastereomers of the group members, aredisclosed separately. When a Markush group or other grouping is usedherein, all individual members of the group and all combinations andsubcombinations possible of the group are intended to be individuallyincluded in the disclosure. When a compound is described herein suchthat a particular isomer, enantiomer or diastereomer of the compound isnot specified, for example, in a formula or in a chemical name, thatdescription is intended to include each isomers and enantiomer of thecompound described individual or in any combination. Additionally,unless otherwise specified, all isotopic variants of compounds disclosedherein are intended to be encompassed by the disclosure. For example, itwill be understood that any one or more hydrogens in a moleculedisclosed can be replaced with deuterium or tritium. Isotopic variantsof a molecule are generally useful as standards in assays for themolecule and in chemical and biological research related to the moleculeor its use. Methods for making such isotopic variants are known in theart. Specific names of compounds are intended to be exemplary, as it isknown that one of ordinary skill in the art can name the same compoundsdifferently.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural reference unless thecontext clearly dictates otherwise. Thus, for example, reference to “acell” includes a plurality of such cells and equivalents thereof knownto those skilled in the art, and so forth. As well, the terms “a” (or“an”), “one or more” and “at least one” can be used interchangeablyherein. It is also to be noted that the terms “comprising”, “including”,and “having” can be used interchangeably. The expression “of any ofclaims XX-YY” (wherein XX and YY refer to claim numbers) is intended toprovide a multiple dependent claim in the alternative form, and in someembodiments is interchangeable with the expression “as in any one ofclaims XX-YY.”

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of ordinary skillin the art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are now described. Nothing herein is to be construed as anadmission that the invention is not entitled to antedate such disclosureby virtue of prior invention.

Whenever a range is given in the specification, for example, a range ofintegers, a temperature range, a time range, a composition range, orconcentration range, all intermediate ranges and subranges, as well asall individual values included in the ranges given are intended to beincluded in the disclosure. As used herein, ranges specifically includethe values provided as endpoint values of the range. As used herein,ranges specifically include all the integer values of the range. Forexample, a range of 1 to 100 specifically includes the end point valuesof 1 and 100. It will be understood that any subranges or individualvalues in a range or subrange that are included in the descriptionherein can be excluded from the claims herein.

As used herein, “comprising” is synonymous and can be usedinterchangeably with “including,” “containing,” or “characterized by,”and is inclusive or open-ended and does not exclude additional,unrecited elements or method steps. As used herein, “consisting of”excludes any element, step, or ingredient not specified in the claimelement. As used herein, “consisting essentially of” does not excludematerials or steps that do not materially affect the basic and novelcharacteristics of the claim. In each instance herein any of the terms“comprising”, “consisting essentially of” and “consisting of” can bereplaced with either of the other two terms. The inventionillustratively described herein suitably can be practiced in the absenceof any element or elements, limitation or limitations which is notspecifically disclosed herein.

One of ordinary skill in the art will appreciate that startingmaterials, biological materials, reagents, synthetic methods,purification methods, analytical methods, assay methods, and biologicalmethods other than those specifically exemplified can be employed in thepractice of the invention without resort to undue experimentation. Allart-known functional equivalents, of any such materials and methods areintended to be included in this invention. The terms and expressionswhich have been employed are used as terms of description and not oflimitation, and there is no intention that in the use of such terms andexpressions of excluding any equivalents of the features shown anddescribed or portions thereof, but it is recognized that variousmodifications are possible within the scope of the invention claimed.Thus, it should be understood that although the invention has beenspecifically disclosed by preferred embodiments and optional features,modification and variation of the concepts herein disclosed can beresorted to by those skilled in the art, and that such modifications andvariations are considered to be within the scope of this invention asdefined by the appended claims.

REFERENCES

-   (1) Momma, T; Hamblin, M. R.; Wu, H. C.; Hasan, T; “Photodynamic    Therapy of Orthotropic Prostate Cancer with Benzoporphyrin    Derivative: Local Control and Distant Metastasis”, Cancer Research,    58, 5425-5431 (December 1998).-   (2) A mechanism-based combination therapy reduces local tumor growth    and metastasis in an orthotopic model of prostate cancer.    Kosharskyy, Boleslav; Solban, Nicolas; Chang, Sung K.; Rizvi, Imran;    Chang, Yuchiao; Hasan, Tayyaba. Department of Anesthesiology, Mount    Sinai Hospital, New York, N.Y., USA. Cancer Research (2006), 66(22),    10953-10958.-   (3) Mechanistic Investigation and Implications of Photodynamic    Therapy Induction of Vascular Endothelial Growth Factor in Prostate    Cancer. Solban, Nicolas; Pal, Selbo K.; Alok, Sinha K.; Sung, Chang    K.; Hasan, Tayyaba. Wellman Center for Photomedicine, Massachusetts    General Hospital, Harvard Medical School, Boston, Mass., USA. Cancer    Research (2006), 66(11), 5633-5640,-   (4) Ferrario A, Gomer C J. Avastin enhances photodynamic therapy    treatment of Kaposi's sarcoma in a mouse tumor model. J Environ    Pathol Toxicol Oncol 2006; 25:251-60.-   (5) Singleton, P. A.; Lingen, M. W.; Fekete, M. J.; Garcia, J. G.    N.; Moss, J. Methylnaltrexone inhibits opiate and VEGF-induced    angiogenesis: role of receptor transactivation. Microvascular    Research (2006), 72(1-2), 3-11.-   (6) Moss, Jonathan; Lingen, Mark; Singleton, Patrick A.; Garcia,    Joe G. N.; Yuan, Chun-Su. Use of opioid antagonists to attenuate    endothelial cell proliferation and migration. U.S. Pat. Appl. Publ.    (2006), 50 pp., Cont.-in-part of Appl. No. PCT/US06/07892. CODEN:    USXXCO US 2006258696 A1 20061116.-   (7) Singleton, Patrick A.; Moreno-Vinasco, Liliana; Sammani, Saad;    Wanderling, Sherry L.; Moss, Jonathan; Garcia, Joe G. N. Attenuation    of vascular permeability by methylnaltrexone: role of mOP-R and S1P3    transactivation. American Journal of Respiratory Cell and Molecular    Biology (2007), 37(2), 222-231. CODEN: AJRBEL ISSN:1044-1549.-   (8) Synergistic effects of methylnaltrexone with 5-fluorouracil and    bevacizumab on inhibition of vascular endothelial growth    factor-induced angiogenesis. Singleton, Patrick A.; Garcia, Joe G.    N.; Moss, Jonathan. Departments of Medicine and Anesthesia and    Critical Care, University of Chicago, Chicago, Ill., USA. Molecular    Cancer Therapeutics (2008), 7(6), 1669-1679. Publisher: American    Association for Cancer Research.-   (9) Lin, Shankung; Tsai, Shiow-Chwen; Lee, Chun-Chung; Wang,    Bao-Wei; Liou, Jer-Young; Shyu, Kou-Gi. Berberine inhibits HIF-1α    expression via enhanced proteolysis. Molecular Pharmacology (2004),    66(3), 612-619.-   (10) Augustin, A J; Puls, S; Offerman I. Triple Therapy for    Choroidal Neovascularization due to Age-Related Macular    Degeneration. Vertportfrin PDT, Bevacizumab, and Dexamethasone.    RETINA 27:133-140, 2007.

What is claimed is:
 1. A method for treating cancer, the methodcomprising; administering to a patient in need of treatment an effectiveamount of a purified (+) enantiomer compound substantially free of acorresponding (−) enantiomer; the (+) enantiomer compound having any ofthe formula:


2. The method of claim 1 comprising a method for treating pancreaticcancer.
 3. The method of claim 1 comprising a method of suppressingtumor growth or tumor metastasis.
 4. The method of claim 1, furthercomprising administering to said patient one or more chemotherapeuticagents.
 5. The method of claim 1, further comprising administering tosaid patient one or more VEGF inhibitors.
 6. The method of claim 1,further comprising administering to said patient 5-fluorouracil,floxuridine, furtulon, capecitabine, gemcitabine, taxol, doxorubicin,cisplatin, thalidomide, paclitaxel, docetaxel or bevacizumab.
 7. Themethod of claim 1, further comprising administering to said patientgemcitabine or bevacizumab.
 8. A method for treating cancer, the methodcomprising: administering to a patient in need of treatment an effectiveamount of a purified (+) enantiomer compound substantially free of acorresponding (−) enantiomer, the (+) enantiomer compound having theformula (FX79) or (FX80):


9. The method of claim 8 comprising a method for treating pancreaticcancer.
 10. The method of claim 8 comprising a method of suppressingtumor growth or tumor metastasis.
 11. The method of claim 8, furthercomprising administering to said patient one or more chemotherapeuticagents.
 12. The method of claim 8, further comprising administering tosaid patient one or more VEGF inhibitors.
 13. The method of claim 8,further comprising administering to said patient 5-fluorouracil,floxuridine, furtulon, capecitabine, gemcitabine, taxol, doxorubicin,cisplatin, thalidomide, paclitaxel, docetaxel or bevacizumab.
 14. Themethod of claim 8, further comprising administering to said patientgemcitabine or bevacizumab.